Molecular Epidemiology of Tuberculosis in Western Burkina Faso: Insights from a MIRU-VNTR Study

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Kaboré, Anselme Millogo, Adama Sourabié, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6636816/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Oct, 2025 Read the published version in BMC Microbiology → Version 1 posted 11 You are reading this latest preprint version Abstract Introduction : Tuberculosis (TB) remains a significant public health challenge in Burkina Faso, with limited molecular epidemiology data available to track strain diversity and transmission patterns. This study aims to characterize the genetic diversity of Mycobacterium tuberculosis complex (MTBC) isolates circulating in Burkina Faso via Mycobacterial Interspersed Repetitive-Unit Variable-Number Tandem-Repeat (MIRU-VNTR) typing. Methods : Genotyping via the MIRU-VNTR technique, which is based on 24 loci, was performed via PCR amplification of DNA extracts from 80 MTBC isolates, followed by agarose gel electrophoresis. The genotypes obtained were identified via comparison with reference strains in the database via the MIRU VNTRplus web application. This study assessed strain diversity, clustering rates, and potential transmission patterns in the study area. Results : Among the 80 isolates, 94.5% were identified as M. tuberculosis sensu stricto, while 2.7% were M. africanum and 2.7% were M. bovis . The predominant lineages were Cameroon (26.3%), Ghana (26.3%), and Haarlem (20%), with the Beijing lineage being rare (1.25%). A low clustering rate of 2.5% was observed. The average discriminating power (allelic diversity index) was 0.444. Eight loci were highly discriminative (h ≥ 0. 6) versus 3 loci that were weakly discriminating (h<0.3). Conclusion : This study provides new insights into the genetic diversity and transmission dynamics of MTBC strains in Burkina Faso. While the observed low clustering suggests limited recent TB transmission, the findings require further validation through larger-scale studies incorporating whole-genome sequencing (WGS). Understanding TB strain diversity is crucial for optimizing diagnostic strategies and TB control policies in West Africa. Molecular epidemiology Mycobacterium tuberculosis complex MIRU-VNTR Tuberculosis transmission Burkina Faso Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION Tuberculosis (TB) remains one of the most prevalent infectious diseases worldwide, with 10.8 million new cases and 1.25 million deaths annually in 2023, according to the World Health Organization (WHO). Despite global TB control efforts, the WHO African Region continues to bear nearly a quarter of the world's new TB cases, with Nigeria and the Democratic Republic of the Congo among the top eight high-burden countries ( 1 ). Burkina Faso, a West African country with approximately 22 million inhabitants, had an estimated TB incidence rate of 45 cases per 100,000 people in 2021. However, 20% of these cases remain bacteriologically unconfirmed, primarily owing to limited diagnostic access and laboratory infrastructure. The true burden of TB transmission and strain diversity in Burkina Faso remain poorly understood, particularly at the molecular level, making epidemiological surveillance a key priority for disease control ( 2 ). Molecular epidemiology plays a critical role in understanding TB transmission patterns, strain diversity, and the effectiveness of control measures. Previous molecular studies in Burkina Faso, including those by Godreuil et al. (2007) ( 3 ) and Gomgnimbou et al. (2012) ( 4 ), identified a TB epidemiological profile dominated by Mycobacterium tuberculosis sensu stricto (MTBss) (80–100%) and M. africanum (0–20%). These studies also reported high clustering rates (~ 80%), indicative of recent active TB transmission in the region( 3 , 4 ). Since these studies, significant changes in the approach to TB control, such as treatment based on the DOTS (directly observed therapy, short-course) strategy and the strengthening of the diagnostic network with the establishment of the GeneXpert® MTB/RIF platforms (Cepheid, USA), have been implemented ( 5 – 7 ). Given these developments, it is crucial to assess whether TB transmission patterns in Burkina Faso have changed and to characterize the genetic diversity of circulating MTBC strains. This study employs Mycobacterial Interspersed Repetitive-Unit Variable-Number Tandem-Repeat (MIRU-VNTR) genotyping, to provide new insights into the molecular epidemiology of TB in Burkina Faso. Materials and Methods Patients, clinical samples, and study area This cross-sectional study was conducted on a retrospective collection of Mycobacterium tuberculosis complex (MTBC) isolates from patients undergoing TB screening or follow-up between February 2013 and December 2016. The samples were collected from two major referral centers in western Burkina Faso, the Regional Center for Tuberculosis Control (RCTC) and the Sourô Sanou University Hospital (SSUH). These centers, located in Bobo-Dioulasso, serve referred patients from the four health regions of western Burkina Faso ( Figure 1 ). According to the Burkina Faso Ministry of Health (2018), these regions account for ~25.7% of the country's TB cases (1,587 out of 6,166 cases), making them important areas for TB molecular epidemiology studies(8). Culture and identification of the M. tuberculosis complex : At the CHUSS Bacteriology Laboratory, sputum samples were decontaminated via Petroff’s method and cultured on Löwenstein–Jensen (LJ) solid media, following Burkina Faso’s National Tuberculosis Control Program guidelines and international protocols (9). Colonies from pure cultures were systematically subjected to Ziehl–Neelsen staining to confirm the presence of acid-fast bacteria before MTBC identification via the SD Bioline TB AgMPT 64 immunochromatographic rapid test (Standard Diagnostics Inc., South Korea) as described elsewhere (10). Rifampicin resistance was determined via the GeneXpert® MTB/RIF platform (Cepheid, USA). Only 80 confirmed MTBC-positive cultures (workflow in Figure 2) were included for further analysis. These isolates were transported to the Bacteriology Laboratory of Arnaud de Villeneuve University Hospital, Montpellier (France), for molecular characterization. Genotyping : Genomic DNA was extracted manually via the GenoLyse® Kit (Hain Life Science GmbH, Nehren, Germany) according to the manufacturer’s protocol (11). MIRU-VNTR typing was performed via a 24-locus MIRU-VNTR panel, with individual amplification of loci via the T100 Thermal Cycler (Bio-Rad, USA), as previously described by Supply et al. (12). The VNTR 4156 locus was inconclusive in some isolates, leading to a 23-locus profile instead of a 24-locus profile. The genotypicprofiles were converted into a numerical 23-locus allelic format for downstream phylogenetic and clustering analysis. Genetic diversity analyses : To explore the genetic variability of the population of isolates studied, the average genetic diversity (H) was calculated using the formula H=S h/n , where h =1- S q i 2 , n=number of loci analyzed, h = allelic diversity of each locus, and the q i 2 relative frequency of the i th allele for the locus considered. The Hunter‒Gaston discriminative index (HGDI) was used to estimate the discriminatory power of MIRU-VNTR. The HGDI was calculated according to the following formula, where ‘’ N’’ is the total number of strains in the typing scheme, ‘’s’’ is the total number of different MIRU-VNTR models, and ‘’ nj’’ is the number of strains belonging to the ‘’jth’’ model (13). Transmission Clustering Analysis. A cluster was defined as two or more isolates with identical MIRU-VNTR genotypes, suggesting potential recent transmission events. The recent transmission rate (RTT) was estimated using the "n-1 method": These rates provide insights into whether transmission is ongoing or whether cases result from past infections or reactivation (14,15). Phylogenetic and Lineage Determination To assess strain relatedness, dendrograms and phylogenetic trees were constructed via UPGMA (unweighted pair group method with arithmetic mean) in PAUP 4.0 (phylogenetic analysis using parsimony) and PHYLIP v3.5c (J. Felsenstein, Seattle, 1993). The resulting phylogenetic trees were visualized and analyzed using Treedyn software. (www.treedyn.org) (16). Lineage classification was performed via the MIRU-VNTRplus database (http://www.miru-vntrplus.org), which compares study strains to 167 globally recognized reference genotypes. A match was considered valid when the genotypic distance between strains was ≤ 0.17. Statistical analysis: Population characteristics were compared via chi-square (χ²) tests. A p value ≤ 0.05 was considered statistically significant. Ethical and deontological aspects This study was approved by the Burkina Faso Ethics Committee for Health Research (Approval No: 2011-8-47). Clinical trial number is not applicable. Written informed consent was obtained from all the participants and guardians of the minors (<18 years old). All patient data were anonymized to ensure confidentiality. RESULTS Characteristics of the population As shown in the flow chart in Figure 2, 92 (26.1%) of the 353 cultured samples were positive, and 13 (3.7%) were contaminated. Among the 80 strains identified as M. tuberculosis complex (MTBC), 64 (80.0%) were new tuberculosis cases from the 303 presumptive tuberculosis samples. These MTBC strains included 61 (76.2%) rifampicin-susceptible strains and 19 (23.8%) rifampicin-resistant strains, according to GeneXpert testing (Cepheid, USA). The strains were obtained from a predominantly male population (sex ratio of 1.42) with a mean age of 32 years (ranging from 8 months to 80 years), 75.35% of patients aged over 16 years, and 33.75% of patients living with HIV. Allelic richness, diversity, and discriminatory power of MIRU-VNTR loci The QUB11B and MIRU40 loci presented the greatest number of alleles, with 8 alleles per locus, and the MIRU2 and MIRU39 loci presented the lowest number of alleles, with 2 alleles per locus. The average discriminating power (allelic diversity index) was 0.444 for the general population. According to this index, 8 loci (ETR A, ETR C, MIRU 26, 40, 52, QUB 11B, 26, VNTR 24) were highly discriminative (h ≥ 0. 6) versus 3 loci (ETRB, MIRU2, MIRU4) that were weakly discriminating (h<0.3). The allelic richness estimated considering the lowest population size varied from 1.95-7.02 and did not differ significantly ( p =0.16) in rifampicin sensitivity. Table 1 shows these different population characteristics according to rifampicin sensitivity. Table 1: Allele number, genetic diversity and allelic richness per locus and per population Number of alleles Genetic diversity Allelic richness * Locus Rif R Rif S General population Rif R Rif S Rif R Rif S ETR A 3 7 7 0.678 0.589 5.263 4.907 ERT B 1 4 5 0 0.127 2.836 3.505 ETR C 4 4 4 0.708 0.683 3.954 3.981 MIRU 2 2 2 2 0.281 0.125 1.955 1.981 MIRU 4 1 3 3 0 0.185 2.735 2.558 MIRU 10 1 5 5 0 0.484 4.658 4.404 MIRU 16 2 3 3 0.199 0.357 2.899 2.936 MIRU 20 2 4 4 0.526 0.501 3.519 3.386 MIRU 23 3 4 4 0.374 0.456 3.879 3.789 MIRU 24 1 3 3 0 0.378 2.899 2.808 MIRU 26 3 6 6 0.561 0.656 4.582 4.258 MIRU 27 1 3 3 0 0.316 2.951 2.873 MIRU 31 3 4 4 0.433 0.496 3.934 3.858 MIRU 39 2 2 2 0.351 0.207 1.996 1.998 MIRU 40 4 8 8 0.573 0.782 7.023 6.63 MIRU 46 2 3 4 0.491 0.577 2.991 3.965 MIRU 49 2 3 3 0.526 0.555 2.998 3 MIRU 52 2 7 7 0.351 0.663 6.106 5.699 QUB 11B 5 8 8 0.713 0.813 6.579 6.49 QUB 26 4 4 7 0.784 0.213 3.035 6.096 VNTR 19 3 4 5 0.368 0.563 3.526 3.829 VNTR 24 3 3 4 0.602 0.67 3 3.809 VNTR 42 3 5 6 0.368 0.242 3.563 3.669 Moyenne 0.386 0.462 3.777 3.932 p value 0.053 0.161 * Allelic richness according to the population with the smallest size N=19 Phylogenetic tree and clustering The phylogenetic tree presented in Figure 3 shows that two (02) strains (2.5%) were grouped in a cluster, whereas the remaining 78 strains (97.5%) were unique individuals. These two strains elonging to the cluster were susceptible to rifampicin. The clustered strain rate and the recent transmission rate were 2.5% and 1.25%, respectively. Identification of the different species affiliated with the strains The identification from the MIRU/VNTRplus online database classified 73 (91.2%) strains into three species and eight different families ( Figure 4 ). Among the 73 identified strains, 3 (3.75%) strains matched with a phylogenetic distance of 0.1364 to the 167 reference strains already described in this database, as summarized in Table 2 . The 73 MTBC strains identified via this platform were distributed between the species MTBss with 69 (94.5%) strains, and the species M. africanum and M. bovis, with 2 (2.7%) strains each. Table 2: Matching of the study strains with the reference strains of the MIRU-VNTRplus database ID ID Reference strain Distance Species Family MLVA MtbC15-9 SpolDB4 ST Rif 33 10476/01 0.1364 M. africanum West Africa2 42-2 326 Se 42 10493/01 0.1364 M. tuberculosis Ghana 68-25 53 Se 56 10439/01/ 10445/01 0.1364 M. tuberculosis Cameroon 73-26 81-26 61 Se Rif : rifampicin; Se : sensitivity; MLVA : multilocus variable number tandem repeat analysis; MtbC15-9 : International nomenclature of MIRU-VNTR genotypes combining the 15 most discriminating loci and 9 auxiliary loci ; SpolDB4 ST : fourth international spoligotyping database shared-types Identification of the families belonging to the strains The Ghana and Cameroon families were the most represented, with 21 (26.3%) strains each, followed by the Haarlem family, with 16 strains each (20.0%). The Beijing family was the least represented, with 1 (1.25%) strain, followed by the Bovis, Uganda II, and West African II families, with 2 (2.5%) strains each, as described in Figures 3&4 . The two strains in the cluster were from the Haarlem family. All resistance strains belonged to the MTBss and spread across all families in varying proportions, except for the two families in Cameroon and Uganda. All strains with unidentified families were resistant (Figure 5). DISCUSSION This study provides a molecular epidemiological assessment of tuberculosis (TB) in western Burkina Faso, investigating strain diversity and transmission patterns via MIRU-VNTR genotyping. More than a decade after the work of Godreuil et al. (2007) ( 3 ), which established the baseline molecular epidemiology of TB in the region, this study re-examines the genotypic landscape of the Mycobacterium tuberculosis complex (MTBC) in the context of ongoing TB control efforts. Given the expansion of GeneXpert® MTB/RIF diagnostics and directly observed therapy (DOTS) strategies, understanding whether these interventions have influenced TB transmission dynamics is crucial for refining public health strategies. Transmission Patterns and Clustering Analysis The study revealed a low clustering rate (2.5%) and a recent transmission rate of 1.25%, which are significantly lower than those reported in Burkina Faso (~ 53.33%) ( 3 ) and West Africa (~ 20%)( 17 ) but closer to the 8% reported in Morocco ( 18 ). Given that a high clustering rate is generally associated with ongoing TB transmission, the decline observed in this study could suggest a shift toward reactivation of latent TB infections rather than recent person‒person transmission. However, the interpretation of these results must be approached with caution. Previous molecular epidemiology studies in Burkina Faso have consistently shown high clustering rates indicative of active transmission. The unexpectedly low clustering observed here suggests either a genuine reduction in TB transmission due to effective TB control efforts or a methodological limitation arising from insufficient sampling and genotyping resolution. Regarding the first explanatory hypothesis, the lower clustering rate may suggest that the TB control interventions introduced over the last decade—including the expansion of GeneXpert® MTB/RIF testing, improved case detection, and DOTS therapy—may have reduced the spread of new infections. A recent study in Burkina Faso suggested that latent TB infection is highly prevalent (~ 87%), with some at-risk groups, such as abattoir workers, reaching 100% latent TB prevalence ( 19 ). The predominance of latent TB activation over new infections could explain the observed low clustering rate. Additionally, the high HIV coinfection rate (33.75%) in this study population suggests that HIV-associated immunosuppression may drive the reactivation of latent TB cases rather than new transmission routes. Nevertheless, another more probable explanation is that the low clustering rate may be an artifact of the study design rather than a true epidemiological phenomenon. Molecular epidemiology studies generally require at least 5–10% case coverage to reliably detect transmission clusters ( 15 ). This study covered only 3.9% of the estimated TB cases in the region, potentially leading to an underrepresentation of microoutbreaks and an overestimation of unique strains. Additionally, MIRU-VNTR genotyping, while effective for strain differentiation, lacks the resolution of whole-genome sequencing (WGS), which can identify microevolutionary changes and closely related isolates that share nearly identical genotypes but differ at the SNP level ( 20 – 22 ). These factors underscore the need for larger-scale studies incorporating WGS to provide a higher-resolution view of TB transmission dynamics. MTBC Lineage Distribution and Global Comparisons: The Cameroon and Ghana lineages were the most prevalent (26.3% each), followed by Haarlem (20%). These findings align with previous reports indicating that Cameroon and Ghana strains are dominant in Burkina Faso and neighboring countries ( 3 , 4 , 23 , 24 ). Additionally, the identification of Ghana and Cameroon strains in international TB databases highlights the potential for cross-border transmission within West Africa and beyond. The low prevalence of the Beijing lineage (1.25%) is also consistent with trends observed in West Africa, where Beijing strains remain rare compared with their higher prevalence in East Asia and parts of Southern Africa ( 17 ). At the species level, the typing strains were distributed predominantly between the species MTBss (94.5%) and marginally between the species of M. africanum and M. bovis (2.7%). Interestingly, the presence of M. bovis (2.7%) was even lower, suggesting that zoonotic tuberculosis remains a concern in Burkina Faso, particularly given the country’s strong reliance on cattle farming. This finding, which aligns with those of previous studies( 3 , 17 , 24 – 26 ), reinforces the need for zoonotic TB surveillance programs to monitor potential animal-to-human transmission. Additionally, the prevalence of M. bovis may be underestimated because of several factors. The first explanation could be the pulmonary origin of the isolates, as M. bovis is preferentially found in extrapulmonary tuberculosis ( 27 ). In addition, the fact that LJ medium enriched with glycerol inhibits some strains of M. bovis can also be considered( 28 , 29 ). Another discrepancy compared with the literature is the low prevalence of M. africanum . This low representation of M. africanum is in agreement with the findings of Godreuil et al. ( 3 ), who reported the exclusive circulation of MTBss. In contrast, Gomgnimbou et al. ( 4 ) reported a significant proportion of M. africanum (22%) alongside a predominant proportion of MTBss (76.4%). This variation in the prevalence of M. africanum can be attributed to the limitations of the techniques used. Indeed, Gomgnimbou et al. ( 4 ) suggested that M. africanum is more fastidious in a subculture (needs pyruvate supplementation to LJ medium) and that the use of DNA extraction from stained slides with Zhiel Nielsen can help overcome this limitation in molecular studies. In addition, M. africanum is more sensitive to first-line antituberculosis drugs, which may explain its low prevalence in populations of patients already treated ( 30 – 32 ). The low proportions of M. africanum reported by Mekonnen et al. raise the question of whether technical limitations are only responsible. In a review conducted in 2019, Mekonnen et al. described an epidemiological profile of tuberculosis in Africa that is largely dominated by MTBss, with only 7% of the isolates being M. africanum ( 17 ). While a certain decrease in the prevalence of M. africanum may have occurred, the factors contributing to this reduction are not well understood. One hypothesis that has been proposed as a partial explanation is the adaptability of species due to evolution. M. africanum is a specialized and ancient pathogen that is geographically limited to West Africa, and it is currently competing with the different so-called modern families of the MTBss species ( 33 , 34 ). Overall, factors contributing to the adaptability of MTBC species, lineages, and families to a particular population or area are not well understood, but some studies suggest that mass BCG vaccination, which has been strictly enforced for decades in this region, may have significantly altered the population structure of MTBC( 35 , 36 ). The predominance of the Cameroon family may reflect historical and ongoing intraregional movement facilitated by the establishment of the Economic Community of West African States (ECOWAS) in 1979. Interestingly, this lineage has also been isolated in France and Belgium, suggesting migration from West Africa to Europe ( 37 ). Rifampicin Resistance and Implications for Drug-Resistant TB Control: In terms of resistance patterns, the proportion of rifampicin-resistant TB cases (23.8%) detected in this study is consistent with regional estimates for West Africa, where multidrug-resistant TB (MDR-TB) remains a growing concern ( 38 ). While GeneXpert® MTB/RIF provides a rapid and accessible diagnostic tool, it has limitations in detecting specific resistance mutations in genes such as rpoB, katG, and inhA. This limitation underscores the need for complementary molecular techniques, such as Line Probe Assays (LPA) or WGS, to provide a more comprehensive understanding of TB drug resistance profiles. Additionally, 33.75% of the TB patients in this study were HIV positive, further emphasizing the need for integrated TB-HIV surveillance and management programs. The high prevalence of HIV-TB coinfection in Burkina Faso suggests that targeted screening and preventive therapy strategies should be reinforced, particularly among high-risk populations. Study Limitations and Future Directions: While this study provides valuable molecular epidemiological data, several limitations must be acknowledged. First, the small sample size (n = 80) limits the ability to detect robust transmission trends, and larger-scale studies with 200 or more isolates would provide more representative epidemiological insights. Second, MIRU-VNTR genotyping, while useful, lacks the resolution of WGS, meaning that some genetically related strains may have been classified as unique owing to the lower discriminatory power of the method. Future studies should integrate WGS-based SNP analysis to delineate better transmission networks and microevolutionary changes within the MTBC population. Furthermore, this study did not include clinical or contact-tracing data, which limits the ability to correlate molecular findings with patient-level transmission risk factors. Future studies should incorporate detailed patient demographics, TB history, treatment adherence data, and epidemiological linkages to strengthen public health applications of molecular surveillance. Finally, expanding drug resistance profiling to include mutations in katG and inhA would provide a more comprehensive picture of drug-resistant TB evolution in the region. Conclusion This study contributes new insights into the genetic diversity and transmission patterns of TB in western Burkina Faso, highlighting the predominance of Cameroon, Ghana, and Haarlem lineages, the low clustering rate, and the significant proportion of rifampicin-resistant strains. While the observed low clustering suggests limited recent TB transmission, the findings require further validation through larger-scale studies incorporating WGS and epidemiological data. From a public health perspective, strengthening TB molecular surveillance and expanding drug resistance monitoring remain key priorities for TB control in Burkina Faso. Future efforts should focus on scaling genotyping approaches, integrating epidemiological data with molecular findings, and reinforcing TB-HIV collaborative interventions. Additionally, enhancing cross-border surveillance for highly prevalent TB lineages such as Cameroon and Ghana strains will be essential in controlling TB transmission within West Africa and beyond. Declarations Ethics approval and consent to participate: This study was approved by the Burkina Faso Ethics Committee for Health Research (Approval No: 2011-8-47). Written informed consent was obtained from all the participants and guardians of the minors (<18 years old). The study adhered to the Declaration of Helsinki, and all patient data were anonymized to ensure confidentiality. Consent for publication: Not applicable Availability of data and materials: The full-length gel DNA electrophoresis images used and/or analysed during the current study are available as supplementary material. Competing interests: The authors declare that they have no competing interests Funding: This study was supported by the Burkina Faso Government and French Embassy bursary Author contributions: A.S.O and S.G: Initiation, design and scientific guarantor of the study; A.M: collection of the samples and preliminary analysis; EB, AP, AS: Patient recrutement and clinical investigation; A.N: molecular analysis, data analysis, bibliographic research and writing of the first manuscript; E.Z, Y.S: Prepared the figures; C.C: Supervision of molecular analysis; O.K, A.S, C.A.O, J.K, A.M.G.B, J.H: Rechecking of the tables, first reading and reorientation of the manuscript; all authors read and approved the final version of the manuscript. Acknowledgments: The authors would like to thank all the technical staff of the CHUSS and Arnaud de Ville Neuve bacteriology laboratories for their contribution to the analysis of the study samples. References World Health Organization. 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Glob Health Action. 2017;10(1):1399749. Chihota V, Niehaus A, Streicher E, Wang X, Sampson S, Mason P, et al. Geospatial distribution of Mycobacterium tuberculosis genotypes in Africa. PLOS ONE. 2018 Aug 1;13:e0200632. World Health Organization. Global tuberculosis report 2022 [Internet]. World Health Organization; 2022 [cited 2023 Apr 8]. xiii, 51 p. Available from: https://apps.who.int/iris/handle/10665/363752 Additional Declarations No competing interests reported. Supplementary Files SupplementarydataonMTBgenoypage.xls DNAGelelectrophoresisrawimages060625VF.pdf Cite Share Download PDF Status: Published Journal Publication published 28 Oct, 2025 Read the published version in BMC Microbiology → Version 1 posted Editorial decision: Revision requested 01 Aug, 2025 Reviews received at journal 04 Jul, 2025 Reviewers agreed at journal 16 Jun, 2025 Reviewers agreed at journal 13 Jun, 2025 Reviews received at journal 13 Jun, 2025 Reviewers agreed at journal 13 Jun, 2025 Reviewers invited by journal 11 Jun, 2025 Editor assigned by journal 11 Jun, 2025 Editor invited by journal 11 Jun, 2025 Submission checks completed at journal 06 Jun, 2025 First submitted to journal 06 Jun, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6636816","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":470206717,"identity":"c28fa683-8c76-4f92-a238-b7a4520e6181","order_by":0,"name":"André Nagalo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6UlEQVRIiWNgGAWjYLCCByDiAAMjiObhI0pLAkQLswFICxspWtgkQDRBLebsxy9+SKiwy+O7ffhY5dccOxk2BuaHj27g0WLZk1MskXAmuVjyXFrabdltyUCHsRkb5+DRYnAgJ0EisY05ccMZHrPbktuYgVp42KTxajn/JvlHYls9UAv/t2LJbfVEaLmRfgxoy2GQLWyMH7cdJkbLGzaLhDPHE2eeYTOWZtx2nIeNmZBfzqc/vvGhojqx7wzzw48/t1Xb87M3P3yMTwsw7gzgTGYeMIlXOQiwP4AzGX8QVD0KRsEoGAUjEQAA64xMDsxE1d8AAAAASUVORK5CYII=","orcid":"","institution":"Université de Montpellier","correspondingAuthor":true,"prefix":"","firstName":"André","middleName":"","lastName":"Nagalo","suffix":""},{"id":470206720,"identity":"3105d399-96be-45f6-94d4-dbe94943faba","order_by":1,"name":"Odilon D. Kaboré","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Odilon","middleName":"D.","lastName":"Kaboré","suffix":""},{"id":470206727,"identity":"ad219a9b-33bd-4d65-a7c1-0a46047f6c30","order_by":2,"name":"Anselme Millogo","email":"","orcid":"","institution":"CHU Sourô Sanou","correspondingAuthor":false,"prefix":"","firstName":"Anselme","middleName":"","lastName":"Millogo","suffix":""},{"id":470206730,"identity":"8cf2037b-f635-4205-94a1-78720cd47cd3","order_by":3,"name":"Adama Sourabié","email":"","orcid":"","institution":"CHU Sourô Sanou","correspondingAuthor":false,"prefix":"","firstName":"Adama","middleName":"","lastName":"Sourabié","suffix":""},{"id":470206731,"identity":"ec28548b-1991-43b8-b0d4-da5240acc28f","order_by":4,"name":"Emmanuel Zongo","email":"","orcid":"","institution":"Laboratoire des pathogènes émergents et réémergents","correspondingAuthor":false,"prefix":"","firstName":"Emmanuel","middleName":"","lastName":"Zongo","suffix":""},{"id":470206732,"identity":"787fe2c7-7db6-4ba6-8e3a-d1fb59c15bcf","order_by":5,"name":"Cheick Ahmed Ouattara","email":"","orcid":"","institution":"Laboratoire des pathogènes émergents et réémergents","correspondingAuthor":false,"prefix":"","firstName":"Cheick","middleName":"Ahmed","lastName":"Ouattara","suffix":""},{"id":470206733,"identity":"c8e6d936-0d12-4f34-8029-5b64ec581287","order_by":6,"name":"Yacouba Sawadogo","email":"","orcid":"","institution":"Laboratoire des pathogènes émergents et réémergents","correspondingAuthor":false,"prefix":"","firstName":"Yacouba","middleName":"","lastName":"Sawadogo","suffix":""},{"id":470206734,"identity":"219f5af8-cccb-4592-a907-651341edda88","order_by":7,"name":"Jacques Kaboré","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Jacques","middleName":"","lastName":"Kaboré","suffix":""},{"id":470206735,"identity":"5c7d1188-0279-47e7-878f-4a2bea733753","order_by":8,"name":"Emile Birba","email":"","orcid":"","institution":"CHU Sourô Sanou","correspondingAuthor":false,"prefix":"","firstName":"Emile","middleName":"","lastName":"Birba","suffix":""},{"id":470206737,"identity":"5f8385a9-ae19-42c4-826d-8435e7a19a82","order_by":9,"name":"Armel Poda","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Armel","middleName":"","lastName":"Poda","suffix":""},{"id":470206739,"identity":"3b87ea9e-7b1e-4cf7-a9d2-d7feb087459e","order_by":10,"name":"Adrien Marie Gaston Belem","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Adrien","middleName":"Marie Gaston","lastName":"Belem","suffix":""},{"id":470206740,"identity":"21748a23-54c7-4b48-88f8-d96f1ae8401a","order_by":11,"name":"Jonathan Hoffmann","email":"","orcid":"","institution":"Fondation Mérieux","correspondingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"","lastName":"Hoffmann","suffix":""},{"id":470206741,"identity":"950da40c-0b8a-49b2-9d4b-03f42234052b","order_by":12,"name":"Christian Carrière","email":"","orcid":"","institution":"Université de Montpellier","correspondingAuthor":false,"prefix":"","firstName":"Christian","middleName":"","lastName":"Carrière","suffix":""},{"id":470206743,"identity":"df7537a3-b788-46fc-ad23-6f7c27a0975b","order_by":13,"name":"Sylvain Godreuil","email":"","orcid":"","institution":"Université de Montpellier","correspondingAuthor":false,"prefix":"","firstName":"Sylvain","middleName":"","lastName":"Godreuil","suffix":""},{"id":470206744,"identity":"120d2779-e0af-46f9-9755-49aec3c8a7a1","order_by":14,"name":"Abdoul-Salam Ouédraogo","email":"","orcid":"","institution":"Laboratoire des pathogènes émergents et réémergents","correspondingAuthor":false,"prefix":"","firstName":"Abdoul-Salam","middleName":"","lastName":"Ouédraogo","suffix":""}],"badges":[],"createdAt":"2025-05-10 21:38:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6636816/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6636816/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12866-025-04430-5","type":"published","date":"2025-10-28T15:58:44+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":84545050,"identity":"6a221758-7af4-4bab-b50a-f5dae77dfd46","added_by":"auto","created_at":"2025-06-13 09:11:42","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":170560,"visible":true,"origin":"","legend":"\u003cp\u003eMap of Burkina Faso showing the regions in the west of the country whose populations attend the CRLAT and CHUSS (A. NAGALO, 2023)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/28794b6fda21c1964d8e3c43.png"},{"id":84544013,"identity":"8e664694-4fbb-4be4-a7e8-da7d391a27ee","added_by":"auto","created_at":"2025-06-13 09:03:42","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":126800,"visible":true,"origin":"","legend":"\u003cp\u003eFlow chart of the stains characterization process\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eLegend: LJ\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: LÖwenstein-Jensen; \u003c/em\u003e\u003cem\u003e\u003cstrong\u003eNTM\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: Non-Tuberculosis Mycobacteria, \u003c/em\u003e\u003cem\u003e\u003cstrong\u003eMTBC\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: Mycobacterium tuberculosis complex; \u003c/em\u003e\u003cem\u003e\u003cstrong\u003eRif\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: Rifampicin; \u003c/em\u003e\u003cem\u003e\u003cstrong\u003eHIV\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: Human Immunodeficiency Virus; \u003c/em\u003e\u003cem\u003e\u003cstrong\u003eMIRU/VNTR\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e: Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/575369f850b602c7e84f8bb4.png"},{"id":84545052,"identity":"4fc05a3a-5d5f-4ff7-84db-fe9d466221cd","added_by":"auto","created_at":"2025-06-13 09:11:42","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":147426,"visible":true,"origin":"","legend":"\u003cp\u003eUPGMA tree based on MIRU-VNTR\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/e75b3ecba2413b396f1b6969.png"},{"id":84545051,"identity":"82b723d7-5edb-42c8-9607-504e7c4ed28a","added_by":"auto","created_at":"2025-06-13 09:11:42","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":83995,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution of strains by family\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/ca9df4901952f6804b217dad.png"},{"id":84544018,"identity":"63299616-3a94-4368-9a40-2fb7cd18fbd6","added_by":"auto","created_at":"2025-06-13 09:03:42","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":70868,"visible":true,"origin":"","legend":"\u003cp\u003eReference to rifampicin by lineage and species(sized by count)\u003c/p\u003e\n\u003cp\u003eND: Non-identified lineage\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/7661f0d02603ca9878c218ed.png"},{"id":95040820,"identity":"eb35b0c1-9ffb-4de5-8a24-a6b2efbe2e49","added_by":"auto","created_at":"2025-11-03 16:10:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1808271,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/732a5be1-2404-46dc-8e92-1afb5f2f9338.pdf"},{"id":84545919,"identity":"a72a29ff-5a28-4544-8976-7ecd1531d06f","added_by":"auto","created_at":"2025-06-13 09:19:42","extension":"xls","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":42496,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementarydataonMTBgenoypage.xls","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/39fefc674c3c9ded10c46510.xls"},{"id":84544039,"identity":"ca51f6d8-5e54-4fae-9df4-ca99f5a31989","added_by":"auto","created_at":"2025-06-13 09:03:43","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":27960188,"visible":true,"origin":"","legend":"","description":"","filename":"DNAGelelectrophoresisrawimages060625VF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6636816/v1/2487e435f14265ec770f1c43.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Molecular Epidemiology of Tuberculosis in Western Burkina Faso: Insights from a MIRU-VNTR Study","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eTuberculosis (TB) remains one of the most prevalent infectious diseases worldwide, with 10.8\u0026nbsp;million new cases and 1.25\u0026nbsp;million deaths annually in 2023, according to the World Health Organization (WHO). Despite global TB control efforts, the WHO African Region continues to bear nearly a quarter of the world's new TB cases, with Nigeria and the Democratic Republic of the Congo among the top eight high-burden countries (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). Burkina Faso, a West African country with approximately 22\u0026nbsp;million inhabitants, had an estimated TB incidence rate of 45 cases per 100,000 people in 2021. However, 20% of these cases remain bacteriologically unconfirmed, primarily owing to limited diagnostic access and laboratory infrastructure. The true burden of TB transmission and strain diversity in Burkina Faso remain poorly understood, particularly at the molecular level, making epidemiological surveillance a key priority for disease control (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Molecular epidemiology plays a critical role in understanding TB transmission patterns, strain diversity, and the effectiveness of control measures. Previous molecular studies in Burkina Faso, including those by Godreuil et al. (2007) (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e) and Gomgnimbou et al. (2012) (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e), identified a TB epidemiological profile dominated by \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e sensu stricto (MTBss) (80\u0026ndash;100%) and \u003cem\u003eM. africanum\u003c/em\u003e (0\u0026ndash;20%). These studies also reported high clustering rates (~\u0026thinsp;80%), indicative of recent active TB transmission in the region(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSince these studies, significant changes in the approach to TB control, such as treatment based on the DOTS (directly observed therapy, short-course) strategy and the strengthening of the diagnostic network with the establishment of the GeneXpert\u0026reg; MTB/RIF platforms (Cepheid, USA), have been implemented (\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Given these developments, it is crucial to assess whether TB transmission patterns in Burkina Faso have changed and to characterize the genetic diversity of circulating MTBC strains. This study employs Mycobacterial Interspersed Repetitive-Unit Variable-Number Tandem-Repeat (MIRU-VNTR) genotyping, to provide new insights into the molecular epidemiology of TB in Burkina Faso.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003ePatients, clinical samples, and study area\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis cross-sectional study was conducted on a retrospective collection of \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e complex (MTBC) isolates from patients undergoing TB screening or follow-up between February 2013 and December 2016. The samples were collected from two major referral centers in western Burkina Faso, the Regional Center for Tuberculosis Control (RCTC) and the Sour\u0026ocirc; Sanou University Hospital (SSUH). These centers, located in Bobo-Dioulasso, serve referred patients from the four health regions of western Burkina Faso (\u003cstrong\u003eFigure 1\u003c/strong\u003e). According to the Burkina Faso Ministry of Health (2018), these regions account for ~25.7% of the country\u0026apos;s TB cases (1,587 out of 6,166 cases), making them important areas for TB molecular epidemiology studies(8).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCulture and identification of the \u003cem\u003eM. tuberculosis\u003c/em\u003e complex\u003c/strong\u003e: At the CHUSS Bacteriology Laboratory, sputum samples were decontaminated via Petroff\u0026rsquo;s method and cultured on L\u0026ouml;wenstein\u0026ndash;Jensen (LJ) solid media, following Burkina Faso\u0026rsquo;s National Tuberculosis Control Program guidelines and international protocols (9). Colonies from pure cultures were systematically subjected to Ziehl\u0026ndash;Neelsen staining to confirm the presence of acid-fast bacteria before MTBC identification via the SD Bioline TB AgMPT 64 immunochromatographic rapid test (Standard Diagnostics Inc., South Korea) as described elsewhere (10). Rifampicin resistance was determined via the GeneXpert\u0026reg; MTB/RIF platform (Cepheid, USA). Only 80 confirmed MTBC-positive cultures (workflow in Figure 2) were included for further analysis. These isolates were transported to the Bacteriology Laboratory of Arnaud de Villeneuve University Hospital, Montpellier (France), for molecular characterization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenotyping\u003c/strong\u003e: Genomic DNA was extracted manually via the GenoLyse\u0026reg; Kit (Hain Life Science GmbH, Nehren, Germany) according to the manufacturer\u0026rsquo;s protocol (11). MIRU-VNTR typing was performed via a 24-locus MIRU-VNTR panel, with individual amplification of loci via the T100 Thermal Cycler (Bio-Rad, USA), as previously described by Supply et al. (12). The VNTR 4156 locus was inconclusive in some isolates, leading to a 23-locus profile instead of a 24-locus profile. The genotypicprofiles were converted into a numerical 23-locus allelic format for downstream phylogenetic and clustering analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenetic diversity analyses\u003c/strong\u003e: To explore the genetic variability of the population of isolates studied, the average genetic diversity (H) was calculated using the formula H=S\u003cem\u003eh/n\u003c/em\u003e, where \u003cem\u003eh\u003c/em\u003e=1- S\u003cem\u003eq\u003csub\u003ei\u003c/sub\u003e\u003csup\u003e2\u003c/sup\u003e\u003c/em\u003e, n=number of loci analyzed, \u003cem\u003eh\u003c/em\u003e= allelic diversity of each locus, and the q\u003csub\u003ei\u003c/sub\u003e\u003csup\u003e2\u003c/sup\u003e relative frequency of the i\u003csup\u003eth\u003c/sup\u003e allele for the locus considered. The Hunter‒Gaston discriminative index (HGDI) was used to estimate the discriminatory power of MIRU-VNTR. The HGDI was calculated according to the following formula, where \u0026lsquo;\u0026rsquo;\u003cem\u003eN\u0026rsquo;\u0026rsquo;\u003c/em\u003e is the total number of strains in the typing scheme, \u0026lsquo;\u0026rsquo;s\u0026rsquo;\u0026rsquo; is the total number of different MIRU-VNTR models, and \u0026lsquo;\u0026rsquo;\u003cem\u003enj\u0026rsquo;\u0026rsquo;\u003c/em\u003e is the number of strains belonging to the \u0026lsquo;\u0026rsquo;jth\u0026rsquo;\u0026rsquo; model (13).\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"431\" height=\"130\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTransmission Clustering Analysis.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA cluster was defined as two or more isolates with identical MIRU-VNTR genotypes, suggesting potential recent transmission events. The recent transmission rate (RTT) was estimated using the \u0026quot;n-1 method\u0026quot;:\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"895\" height=\"226\"\u003e\u003c/p\u003e\n\u003cp\u003eThese rates provide insights into whether transmission is ongoing or whether cases result from past infections or reactivation (14,15).\u003c/p\u003e\n\u003ch3 id=\"_Toc76651984\"\u003ePhylogenetic and Lineage Determination\u003c/h3\u003e\n\u003cp\u003eTo assess strain relatedness, dendrograms and phylogenetic trees were constructed via UPGMA (unweighted pair group method with arithmetic mean) in PAUP 4.0 (phylogenetic analysis using parsimony) and PHYLIP v3.5c (J. Felsenstein, Seattle, 1993).\u003c/p\u003e\n\u003cp\u003eThe resulting phylogenetic trees were visualized and analyzed using Treedyn software.\u003c/p\u003e\n\u003cp\u003e(www.treedyn.org) (16).\u003c/p\u003e\n\u003cp\u003eLineage classification was performed via the MIRU-VNTRplus database (http://www.miru-vntrplus.org), which compares study strains to 167 globally recognized reference genotypes. A match was considered valid when the genotypic distance between strains was \u0026le; 0.17.\u003c/p\u003e\n\u003cp id=\"_Toc76651986\"\u003e\u003cstrong\u003eStatistical\u0026nbsp;\u003c/strong\u003eanalysis: Population characteristics were compared via chi-square (\u0026chi;\u0026sup2;) tests. A p value \u0026le; 0.05 was considered statistically significant.\u003c/p\u003e\n\u003ch2\u003eEthical and deontological aspects\u003c/h2\u003e\n\u003cp\u003eThis study was approved by the Burkina Faso Ethics Committee for Health Research (Approval No: 2011-8-47). Clinical trial number is not applicable. Written informed consent was obtained from all the participants and guardians of the minors (\u0026lt;18 years old). All patient data were anonymized to ensure confidentiality.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003e\u003cstrong\u003eCharacteristics of the population\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAs shown in the flow chart in Figure 2, 92 (26.1%) of the 353 cultured samples were positive, and 13 (3.7%) were contaminated. Among the 80 strains identified as \u003cem\u003eM. tuberculosis\u0026nbsp;\u003c/em\u003ecomplex (MTBC), 64 (80.0%) were new tuberculosis cases from the 303 presumptive tuberculosis samples. These MTBC strains included 61 (76.2%) rifampicin-susceptible strains and 19 (23.8%) rifampicin-resistant strains, according to GeneXpert testing (Cepheid, USA). The strains were obtained from a predominantly male population (sex ratio of 1.42) with a mean age of 32 years (ranging from 8 months to 80 years), 75.35% of patients aged over 16 years, and 33.75% of patients living with HIV.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eAllelic richness, diversity, and discriminatory power of MIRU-VNTR loci\u003c/h2\u003e\n\u003cp\u003eThe QUB11B and MIRU40 loci presented the greatest number of alleles, with 8 alleles per locus, and the MIRU2 and MIRU39 loci presented the lowest number of alleles, with 2 alleles per locus.\u003c/p\u003e\n\u003cp\u003eThe average discriminating power (allelic diversity index) was 0.444 for the general population. According to this index, 8 loci (ETR A, ETR C, MIRU 26, 40, 52, QUB 11B, 26, VNTR 24) were highly discriminative (h \u0026ge; 0. 6) versus 3 loci (ETRB, MIRU2, MIRU4) that were weakly discriminating (h\u0026lt;0.3). The allelic richness estimated considering the lowest population size varied from 1.95-7.02 and did not differ significantly (\u003cem\u003ep\u003c/em\u003e=0.16) in rifampicin sensitivity. \u003cstrong\u003eTable 1\u003c/strong\u003e shows these different population characteristics according to rifampicin sensitivity.\u003c/p\u003e\n\u003cp\u003eTable 1: Allele number, genetic diversity and allelic richness per locus and per population\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"686\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"bottom\" style=\"width: 221px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of alleles\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 184px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGenetic diversity\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAllelic richness *\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLocus\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif R\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif S\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeneral population\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif R\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif S\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif R\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif S\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eETR A\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.678\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.589\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e5.263\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e4.907\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eERT B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.836\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.505\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eETR C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.708\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.683\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.954\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.981\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.281\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e1.955\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e1.981\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.185\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.735\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e2.558\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.484\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e4.658\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e4.404\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.199\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.357\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e2.936\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.526\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.501\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.519\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.386\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.374\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.456\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.789\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.378\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e2.808\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.561\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.656\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e4.582\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e4.258\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.951\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e2.873\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.433\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.496\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.934\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.858\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.351\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e1.996\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e1.998\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.573\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.782\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e7.023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e6.63\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.491\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.991\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.965\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.526\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.555\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e2.998\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMIRU 52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.351\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.663\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e6.106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e5.699\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eQUB 11B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.713\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.813\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e6.579\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e6.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eQUB 26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.784\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.213\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.035\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e6.096\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eVNTR 19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.368\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.563\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.526\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.829\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eVNTR 24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.602\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.67\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.809\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eVNTR 42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 89px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e0.368\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e0.242\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e3.563\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e3.669\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMoyenne\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 59px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 89px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.386\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.462\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 111px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.777\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.932\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 59px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 89px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 184px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.053\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.161\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003e*\u003c/em\u003e \u003cem\u003eAllelic richness according to the population with the smallest size N=19\u003c/em\u003e\u003c/p\u003e\n\u003ch2\u003ePhylogenetic tree and clustering\u003c/h2\u003e\n\u003cp\u003eThe phylogenetic tree presented in \u003cstrong\u003eFigure 3\u003c/strong\u003e shows that two (02) strains (2.5%) were grouped in a cluster, whereas the remaining 78 strains (97.5%) were unique individuals.\u003c/p\u003e\n\u003cp\u003eThese two strains elonging to the cluster were susceptible to rifampicin. The clustered strain rate and the recent transmission rate were 2.5% and 1.25%, respectively.\u003c/p\u003e\n\u003ch3\u003eIdentification of the different species affiliated with the strains\u003c/h3\u003e\n\u003cp\u003eThe identification from the MIRU/VNTRplus online database classified 73 (91.2%) strains into three species and eight different families (\u003cstrong\u003eFigure 4\u003c/strong\u003e). Among the 73 identified strains, 3 (3.75%) strains matched with a phylogenetic distance of 0.1364 to the 167 reference strains already described in this database, as summarized in \u003cstrong\u003eTable 2\u003c/strong\u003e. The 73 MTBC strains identified via this platform were distributed between the species MTBss with 69 (94.5%) strains, and the species \u003cem\u003eM. africanum\u003c/em\u003e and \u003cem\u003eM. bovis,\u003c/em\u003e with 2 (2.7%) strains each.\u003c/p\u003e\n\u003cp\u003eTable 2: Matching of the study strains with the reference strains of the MIRU-VNTRplus database\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eID\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eID Reference strain\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDistance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFamily\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMLVA\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eMtbC15-9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpolDB4 ST\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRif\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e10476/01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.1364\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cem\u003eM. africanum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003eWest Africa2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e42-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e326\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eSe\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e10493/01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.1364\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cem\u003eM. tuberculosis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003eGhana\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e68-25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eSe\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e10439/01/\u003c/p\u003e\n \u003cp\u003e10445/01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.1364\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cem\u003eM. tuberculosis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003eCameroon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e73-26\u003c/p\u003e\n \u003cp\u003e81-26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eSe\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eRif\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e: rifampicin; \u003cstrong\u003eSe\u003c/strong\u003e: sensitivity; \u003cstrong\u003eMLVA\u003c/strong\u003e: multilocus variable number tandem repeat analysis;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMtbC15-9\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e: International nomenclature of MIRU-VNTR genotypes combining the 15 most discriminating loci and 9 auxiliary loci\u003cstrong\u003e; SpolDB4 ST\u003c/strong\u003e: fourth international spoligotyping database shared-types\u003c/em\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of the families belonging to the strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Ghana and Cameroon families were the most represented, with 21 (26.3%) strains each, followed by the Haarlem family, with 16 strains each (20.0%). The Beijing family was the least represented, with 1 (1.25%) strain, followed by the Bovis, Uganda II, and West African II families, with 2 (2.5%) strains each, as described in \u003cstrong\u003eFigures 3\u0026amp;4\u003c/strong\u003e. The two strains in the cluster were from the Haarlem family. All resistance strains belonged to the MTBss and spread across all families in varying proportions, except for the two families in Cameroon and Uganda. All strains with unidentified families were resistant (Figure 5).\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThis study provides a molecular epidemiological assessment of tuberculosis (TB) in western Burkina Faso, investigating strain diversity and transmission patterns via MIRU-VNTR genotyping. More than a decade after the work of Godreuil et al. (2007) (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e), which established the baseline molecular epidemiology of TB in the region, this study re-examines the genotypic landscape of the \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e complex (MTBC) in the context of ongoing TB control efforts. Given the expansion of GeneXpert\u0026reg; MTB/RIF diagnostics and directly observed therapy (DOTS) strategies, understanding whether these interventions have influenced TB transmission dynamics is crucial for refining public health strategies.\u003c/p\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eTransmission Patterns and Clustering Analysis\u003c/h2\u003e \u003cp\u003eThe study revealed a low clustering rate (2.5%) and a recent transmission rate of 1.25%, which are significantly lower than those reported in Burkina Faso (~\u0026thinsp;53.33%) (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e) and West Africa (~\u0026thinsp;20%)(\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e) but closer to the 8% reported in Morocco (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e). Given that a high clustering rate is generally associated with ongoing TB transmission, the decline observed in this study could suggest a shift toward reactivation of latent TB infections rather than recent person‒person transmission. However, the interpretation of these results must be approached with caution. Previous molecular epidemiology studies in Burkina Faso have consistently shown high clustering rates indicative of active transmission. The unexpectedly low clustering observed here suggests either a genuine reduction in TB transmission due to effective TB control efforts or a methodological limitation arising from insufficient sampling and genotyping resolution. Regarding the first explanatory hypothesis, the lower clustering rate may suggest that the TB control interventions introduced over the last decade\u0026mdash;including the expansion of GeneXpert\u0026reg; MTB/RIF testing, improved case detection, and DOTS therapy\u0026mdash;may have reduced the spread of new infections. A recent study in Burkina Faso suggested that latent TB infection is highly prevalent (~\u0026thinsp;87%), with some at-risk groups, such as abattoir workers, reaching 100% latent TB prevalence (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). The predominance of latent TB activation over new infections could explain the observed low clustering rate. Additionally, the high HIV coinfection rate (33.75%) in this study population suggests that HIV-associated immunosuppression may drive the reactivation of latent TB cases rather than new transmission routes. Nevertheless, another more probable explanation is that the low clustering rate may be an artifact of the study design rather than a true epidemiological phenomenon. Molecular epidemiology studies generally require at least 5\u0026ndash;10% case coverage to reliably detect transmission clusters (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e). This study covered only 3.9% of the estimated TB cases in the region, potentially leading to an underrepresentation of microoutbreaks and an overestimation of unique strains. Additionally, MIRU-VNTR genotyping, while effective for strain differentiation, lacks the resolution of whole-genome sequencing (WGS), which can identify microevolutionary changes and closely related isolates that share nearly identical genotypes but differ at the SNP level (\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e). These factors underscore the need for larger-scale studies incorporating WGS to provide a higher-resolution view of TB transmission dynamics.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eMTBC Lineage Distribution and Global Comparisons:\u003c/h2\u003e \u003cp\u003eThe Cameroon and Ghana lineages were the most prevalent (26.3% each), followed by Haarlem (20%). These findings align with previous reports indicating that Cameroon and Ghana strains are dominant in Burkina Faso and neighboring countries (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e). Additionally, the identification of Ghana and Cameroon strains in international TB databases highlights the potential for cross-border transmission within West Africa and beyond. The low prevalence of the Beijing lineage (1.25%) is also consistent with trends observed in West Africa, where Beijing strains remain rare compared with their higher prevalence in East Asia and parts of Southern Africa (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAt the species level, the typing strains were distributed predominantly between the species MTBss (94.5%) and marginally between the species of \u003cem\u003eM. africanum\u003c/em\u003e and \u003cem\u003eM. bovis\u003c/em\u003e (2.7%). Interestingly, the presence of \u003cem\u003eM. bovis\u003c/em\u003e (2.7%) was even lower, suggesting that zoonotic tuberculosis remains a concern in Burkina Faso, particularly given the country\u0026rsquo;s strong reliance on cattle farming. This finding, which aligns with those of previous studies(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e), reinforces the need for zoonotic TB surveillance programs to monitor potential animal-to-human transmission. Additionally, the prevalence of \u003cem\u003eM. bovis\u003c/em\u003e may be underestimated because of several factors. The first explanation could be the pulmonary origin of the isolates, as \u003cem\u003eM. bovis\u003c/em\u003e is preferentially found in extrapulmonary tuberculosis (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). In addition, the fact that LJ medium enriched with glycerol inhibits some strains of \u003cem\u003eM. bovis\u003c/em\u003e can also be considered(\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAnother discrepancy compared with the literature is the low prevalence of \u003cem\u003eM. africanum\u003c/em\u003e. This low representation of \u003cem\u003eM. africanum\u003c/em\u003e is in agreement with the findings of Godreuil et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e), who reported the exclusive circulation of MTBss. In contrast, Gomgnimbou et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e) reported a significant proportion of \u003cem\u003eM. africanum\u003c/em\u003e (22%) alongside a predominant proportion of MTBss (76.4%). This variation in the prevalence of \u003cem\u003eM. africanum\u003c/em\u003e can be attributed to the limitations of the techniques used. Indeed, Gomgnimbou et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e) suggested that \u003cem\u003eM. africanum\u003c/em\u003e is more fastidious in a subculture (needs pyruvate supplementation to LJ medium) and that the use of DNA extraction from stained slides with Zhiel Nielsen can help overcome this limitation in molecular studies. In addition, \u003cem\u003eM. africanum\u003c/em\u003e is more sensitive to first-line antituberculosis drugs, which may explain its low prevalence in populations of patients already treated (\u003cspan additionalcitationids=\"CR31\" citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). The low proportions of \u003cem\u003eM. africanum\u003c/em\u003e reported by Mekonnen et al. raise the question of whether technical limitations are only responsible. In a review conducted in 2019, Mekonnen et al. described an epidemiological profile of tuberculosis in Africa that is largely dominated by MTBss, with only 7% of the isolates being \u003cem\u003eM. africanum\u003c/em\u003e (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). While a certain decrease in the prevalence of \u003cem\u003eM. africanum\u003c/em\u003e may have occurred, the factors contributing to this reduction are not well understood. One hypothesis that has been proposed as a partial explanation is the adaptability of species due to evolution. \u003cem\u003eM. africanum\u003c/em\u003e is a specialized and ancient pathogen that is geographically limited to West Africa, and it is currently competing with the different so-called modern families of the MTBss species (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOverall, factors contributing to the adaptability of MTBC species, lineages, and families to a particular population or area are not well understood, but some studies suggest that mass BCG vaccination, which has been strictly enforced for decades in this region, may have significantly altered the population structure of MTBC(\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e). The predominance of the Cameroon family may reflect historical and ongoing intraregional movement facilitated by the establishment of the Economic Community of West African States (ECOWAS) in 1979. Interestingly, this lineage has also been isolated in France and Belgium, suggesting migration from West Africa to Europe (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eRifampicin Resistance and Implications for Drug-Resistant TB Control:\u003c/h2\u003e \u003cp\u003eIn terms of resistance patterns, the proportion of rifampicin-resistant TB cases (23.8%) detected in this study is consistent with regional estimates for West Africa, where multidrug-resistant TB (MDR-TB) remains a growing concern (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). While GeneXpert\u0026reg; MTB/RIF provides a rapid and accessible diagnostic tool, it has limitations in detecting specific resistance mutations in genes such as rpoB, katG, and inhA. This limitation underscores the need for complementary molecular techniques, such as Line Probe Assays (LPA) or WGS, to provide a more comprehensive understanding of TB drug resistance profiles. Additionally, 33.75% of the TB patients in this study were HIV positive, further emphasizing the need for integrated TB-HIV surveillance and management programs. The high prevalence of HIV-TB coinfection in Burkina Faso suggests that targeted screening and preventive therapy strategies should be reinforced, particularly among high-risk populations.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eStudy Limitations and Future Directions:\u003c/h2\u003e \u003cp\u003eWhile this study provides valuable molecular epidemiological data, several limitations must be acknowledged. First, the small sample size (n\u0026thinsp;=\u0026thinsp;80) limits the ability to detect robust transmission trends, and larger-scale studies with 200 or more isolates would provide more representative epidemiological insights. Second, MIRU-VNTR genotyping, while useful, lacks the resolution of WGS, meaning that some genetically related strains may have been classified as unique owing to the lower discriminatory power of the method. Future studies should integrate WGS-based SNP analysis to delineate better transmission networks and microevolutionary changes within the MTBC population.\u003c/p\u003e \u003cp\u003eFurthermore, this study did not include clinical or contact-tracing data, which limits the ability to correlate molecular findings with patient-level transmission risk factors. Future studies should incorporate detailed patient demographics, TB history, treatment adherence data, and epidemiological linkages to strengthen public health applications of molecular surveillance. Finally, expanding drug resistance profiling to include mutations in katG and inhA would provide a more comprehensive picture of drug-resistant TB evolution in the region.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study contributes new insights into the genetic diversity and transmission patterns of TB in western Burkina Faso, highlighting the predominance of Cameroon, Ghana, and Haarlem lineages, the low clustering rate, and the significant proportion of rifampicin-resistant strains. While the observed low clustering suggests limited recent TB transmission, the findings require further validation through larger-scale studies incorporating WGS and epidemiological data.\u003c/p\u003e \u003cp\u003eFrom a public health perspective, strengthening TB molecular surveillance and expanding drug resistance monitoring remain key priorities for TB control in Burkina Faso. Future efforts should focus on scaling genotyping approaches, integrating epidemiological data with molecular findings, and reinforcing TB-HIV collaborative interventions. Additionally, enhancing cross-border surveillance for highly prevalent TB lineages such as Cameroon and Ghana strains will be essential in controlling TB transmission within West Africa and beyond.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cul\u003e\n \u003cli\u003e\u003cstrong\u003eEthics approval and consent to participate:\u0026nbsp;\u003c/strong\u003eThis study was approved by the Burkina Faso Ethics Committee for Health Research (Approval No: 2011-8-47). Written informed consent was obtained from all the participants and guardians of the minors (\u0026lt;18 years old). The study adhered to the Declaration of Helsinki, and all patient data were anonymized to ensure confidentiality.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eConsent for publication:\u003c/strong\u003e Not applicable\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAvailability of data and materials:\u0026nbsp;\u003c/strong\u003eThe full-length gel DNA electrophoresis images used and/or analysed during the current study are available as supplementary material.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eCompeting interests:\u003c/strong\u003e The authors declare that they have no competing interests\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e This study was supported by the Burkina Faso Government and French Embassy bursary\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAuthor contributions:\u0026nbsp;\u003c/strong\u003eA.S.O and S.G: Initiation, design and scientific guarantor of the study; A.M: collection of the samples and preliminary analysis; \u0026nbsp;EB, AP, AS: Patient recrutement and clinical investigation; A.N: molecular analysis, data analysis, bibliographic research and writing of the first manuscript; E.Z, Y.S: Prepared the figures; C.C: Supervision of molecular analysis; O.K, A.S, C.A.O, J.K, A.M.G.B, J.H: Rechecking of the tables, first reading and reorientation of the manuscript; all authors read and approved the final version of the manuscript.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003eThe authors would like to thank all the technical staff of the CHUSS and Arnaud de Ville Neuve bacteriology laboratories for their contribution to the analysis of the study samples.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eWorld Health Organization. Global Tuberculosis Report 2024. Geneva: World Health Organization; 2024.\u003c/li\u003e\n\u003cli\u003eWorld Health Organization. Tuberculosis profile. 2023 [cited 2023 Apr 8]. Tuberculosis profile. Available from: https://worldhealthorg.shinyapps.io/tb_profiles/?_inputs_\u0026amp;entity_type=%22country%22\u0026amp;lan=%22EN%22\u0026amp;iso2=%22BF%22\u003c/li\u003e\n\u003cli\u003eGodreuil S, Torrea G, Terru D, Chevenet F, Diagbouga S, Supply P, et al. First Molecular Epidemiology Study of Mycobacterium tuberculosis in Burkina Faso. J Clin Microbiol. 2007 Mar 1;45(3):921\u0026ndash;7.\u003c/li\u003e\n\u003cli\u003eGomgnimbou MK, Refr\u0026eacute;gier G, Diagbouga SP, Adama S, Kabor\u0026eacute; A, Ouiminga A, et al. Spoligotyping of \u003cem\u003eMycobacterium africanum\u003c/em\u003e , Burkina Faso. Emerg Infect Dis [Internet]. 2012 Jan [cited 2021 Apr 17];18(1):117\u0026ndash;9. 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Available from: https://apps.who.int/iris/handle/10665/363752\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Molecular epidemiology, Mycobacterium tuberculosis complex, MIRU-VNTR, Tuberculosis transmission, Burkina Faso","lastPublishedDoi":"10.21203/rs.3.rs-6636816/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6636816/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eIntroduction\u003c/strong\u003e: Tuberculosis (TB) remains a significant public health challenge in Burkina Faso, with limited molecular epidemiology data available to track strain diversity and transmission patterns. This study aims to characterize the genetic diversity of \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e complex (MTBC) isolates circulating in Burkina Faso via Mycobacterial Interspersed Repetitive-Unit Variable-Number Tandem-Repeat (MIRU-VNTR) typing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: Genotyping via the MIRU-VNTR technique, which is based on 24 loci, was performed via PCR amplification of DNA extracts from 80 MTBC\u003cem\u003e \u003c/em\u003eisolates, followed by agarose gel electrophoresis. The genotypes obtained were identified via comparison with reference strains in the database via the MIRU VNTRplus web application. This study assessed strain diversity, clustering rates, and potential transmission patterns in the study area.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: Among the 80 isolates, 94.5% were identified as \u003cem\u003eM. tuberculosis\u003c/em\u003e sensu stricto, while 2.7% were \u003cem\u003eM. africanum\u003c/em\u003e and 2.7% were \u003cem\u003eM. bovis\u003c/em\u003e. The predominant lineages were Cameroon (26.3%), Ghana (26.3%), and Haarlem (20%), with the Beijing lineage being rare (1.25%). A low clustering rate of 2.5% was observed. The average discriminating power (allelic diversity index) was 0.444. Eight loci were highly discriminative (h ≥ 0. 6) versus 3 loci that were weakly discriminating (h\u0026lt;0.3).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: This study provides new insights into the genetic diversity and transmission dynamics of MTBC strains in Burkina Faso. While the observed low clustering suggests limited recent TB transmission, the findings require further validation through larger-scale studies incorporating whole-genome sequencing (WGS). Understanding TB strain diversity is crucial for optimizing diagnostic strategies and TB control policies in West Africa.\u003c/p\u003e","manuscriptTitle":"Molecular Epidemiology of Tuberculosis in Western Burkina Faso: Insights from a MIRU-VNTR Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-13 09:03:37","doi":"10.21203/rs.3.rs-6636816/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-08-01T17:54:38+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-04T21:22:40+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"263477286325751303935589537504560846590","date":"2025-06-16T15:47:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"220667070442970715470365644585900058105","date":"2025-06-13T13:52:56+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-06-13T12:36:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"172268926948483418026410347230712902450","date":"2025-06-13T10:44:44+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-06-11T13:45:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-06-11T13:24:16+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-06-11T05:16:08+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-06-07T00:43:08+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Microbiology","date":"2025-06-07T00:40:27+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b5914469-f540-4697-8c7c-3e9d4faec5ed","owner":[],"postedDate":"June 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-11-03T16:08:03+00:00","versionOfRecord":{"articleIdentity":"rs-6636816","link":"https://doi.org/10.1186/s12866-025-04430-5","journal":{"identity":"bmc-microbiology","isVorOnly":false,"title":"BMC Microbiology"},"publishedOn":"2025-10-28 15:58:44","publishedOnDateReadable":"October 28th, 2025"},"versionCreatedAt":"2025-06-13 09:03:37","video":"","vorDoi":"10.1186/s12866-025-04430-5","vorDoiUrl":"https://doi.org/10.1186/s12866-025-04430-5","workflowStages":[]},"version":"v1","identity":"rs-6636816","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6636816","identity":"rs-6636816","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}