Importance of molecular tools in arbovirus and malaria disease coinfection detection in humans, Bobo- Dioulasso, western Burkina Faso

Importance of molecular tools in arbovirus and malaria disease coinfection detection in humans, Bobo- Dioulasso, western Burkina Faso | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Importance of molecular tools in arbovirus and malaria disease coinfection detection in humans, Bobo- Dioulasso, western Burkina Faso Louis Robert Wendyam Belem, Raymond Karlhis Yao, Miriam Félicité Amara, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7658090/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In tropical regions, arbovirus disease and malaria co-circulate currently; consequently, co-infection of both diseases can be found and complicates the diagnosis and treatment process with potentially high morbidity and mortality. This study was designed to demonstrate the co-circulation of arbovirus infection and malaria in Bobo-Dioulasso, as well as the importance of molecular tools in the early detection of coinfection. This cross-sectional study was conducted in Bobo-Dioulasso, Burkina Faso, between June 2023 and August 2023. Participants were included based on clinical symptoms, and blood samples were collected for dengue rapid diagnostic test (RDTs), molecular detection of dengue virus, chikungunya virus, and malaria. Microscopic examination was also performed to diagnose malaria infection. Among 306 samples screened using DENV RT-PCR and Malaria microscopy detection, 5.22% (16/306) were DENV- Plasmodium coinfections. According to DENV screening using RT-PCR and malaria screening using PCR, 7.51% (23/306) were found to be coinfected with DENV and Plasmodium . In this study, 100% (23/23) of the coinfection samples were malaria-positive by PCR, whereas 69.56% (16/23) were positive by microscopy. CHIKV has not been detected in this study. Among coinfections, 74.0% (17/23) were coinfections between DENV-3 and P. falciparum , 13.0% (3/23) between DENV-3 and P. malariae , 8.7% (2/23) between DENV-1 and P. falciparum , and 4.3% (1/23) between DENV-1 and P. malariae. Our study demonstrated the utility of molecular tools in detecting dengue and malaria coinfection in the acute phase. It also showed the co-circulation between DENV-1, DENV-3, P. falciparum, and P. malariae. Molecular arbovirus malaria coinfection Bobo-Dioulasso Figures Figure 1 Figure 2 Figure 3 Introduction Arbovirus infection and malaria are vector-borne diseases and are responsible for major public health issues in the tropical and subtropical regions, including Burkina Faso [ 1 , 2 ]. Dengue virus (DENV) and chikungunya virus (CHIKV) are the main arboviruses in the world and are transmitted mainly by Aedes (Ae) aegypti and Ae. albopictus [ 3 ]. More than 3.9 billion humans in the world are susceptible to contracting DENV, principally in tropical countries [ 4 ]. According to the World Health Organization (WHO), CHIKV is now present in more than 110 countries across Africa, Asia, and the Americas [ 5 ]. CHIKV and DENV are ribonucleic acid (RNA) viruses belonging to the Togaviridae and Flaviviridae families, respectively. DENV is classified into four genetically different serotypes (DENV1-4) [ 6 , 7 ]. Prior infection by one DENV serotype offers long-term immunity to that serotype. However, subsequent infection by a different serotype can cause severe disease like dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) [ 8 ]. Generally, CHIKV infection has low mortality rates. However, it can cause significant morbidity, impacting the quality of life for infected individuals and causing economic losses, mainly in the least developed countries [ 9 ] Malaria is a parasite disease, mainly caused by Plasmodium (P.) falciparum , P. vivax , P. malariae , and P. ovale . The Plasmodium genome is Deoxyribonucleic (DNA) [ 10 ]. In sub-Saharan Africa, P. falciparum is the most common parasite responsible for 90% of the global malaria burden and all malaria deaths[ 11 ]. The Anopheles mosquito transmits malaria, and Anopheles gambiae is considered to be the essential malaria transmission vector in Africa [ 12 ]. It is expected that the highly mobile lifestyle of the population, the increased activities made possible by dependable international transportation networks, and climatic change will increase the prevalence of dengue and malaria co-infection [ 13 ]. Malaria and arbovirus infections presented similar main symptoms, such as fever, headache, myalgia, arthralgia, rash, nausea, diarrhea, vomiting, and abdominal pain [ 14 ]. Malaria is endemic and the first disease responsible for mortality in Burkina Faso. In Burkina, the DENV is the main arbovirus responsible for outbreaks with high prevalence and important morbidity and mortality. Between August and November 2023, Burkina Faso reported important DENV outbreaks and fewer CHIKV outbreaks [ 15 , 16 ]. Several studies have shown that DENV and Malaria can co-circulate [ 1 , 17 , 18 ]. Consequently, the co-circulation of both diseases complicates the diagnosis and treatment process [ 19 ]. In Burkina Faso, many people use antimalarial treatment for a fever or headache without medical consultation and biological diagnosis. This practice can lead to disease complications if the patient is infected by DENV or coinfected with malaria. Although malaria or DENV mono infections can be sometimes severe, coinfections between both diseases could be even more fatal [ 20 ]. Because of their similar clinical manifestations, probable concurrent dengue fever and malaria are often neglected and could lead to misdiagnosis as malaria alone [ 21 ]. During coinfection, misdiagnosis is probably higher than mono-infection; consequently, slow identification of dengue outbreaks can occur and lead to high morbidity or mortality. Serological diagnosis of arbovirus is limited because antibodies such as immunoglobulin M (IgM) and immunoglobulin G (IgG) may not be detectable early in the acute phase of the disease, and cross-reactivity between anti-flavivirus antibodies may occur [ 22 ]. Many commercial rapid diagnostic tests (RDTs) to detect CHIKV and DENV are available [ 23 , 24 ]. However, false-positive test results have been reported in some studies, which is a limitation in the diagnosis [ 25 ]. Several studies have reported that sometimes malaria is misdiagnosed by rapid diagnostic tests (RDTs) due to Histidine-Rich Protein 2 (HRP2) and HRP3 gene deletions [ 26 – 28 ]. Microscopic examination of Giemsa-stained blood slides has been the standard for malaria diagnosis for nearly a century. However, this technique is labor-intensive, time-consuming, and challenged by a high limit of detection (LoD). Under optimal circumstances, the latter is predicted to be nearly 50 parasites/µL and is highly dependent on the slide's quality and the microscopist's level of expertise [ 29 ]. Furthermore, personnel’s lack of practice and proficiency may account for delays and errors in diagnosis. However, molecular methods are the best means of differential diagnosis and can detect coinfection between vector-borne diseases[ 30 ]. In Burkina, DENV and Plasmodium coinfection data are scarce in healthcare and less described due to the diagnostic algorithm shortcoming. This study was designed to demonstrate the importance of molecular tools in the early detection of coinfection, as well as the co-circulation of arbovirus infection, and malaria in Bobo-Dioulasso. Early detection of coinfection between arbovirus and malaria is crucial for appropriate treatment and essential to prevent severe disease and death. Materials and Methods Site and study design This is a cross-sectional study, performed in Bobo-Dioulasso (11° 11′ 00″ North, 4° 17′ 00″ West), belonging to the Guiriko region, Burkina Faso, during the rainy season between June 2023 and August 2023. The study period offers good conditions for mosquito breeding, such as Anopheles and Aedes , that is favorable for malaria and arbovirus transmission. Participants have been included in the medical centers with surgical antenna (CMA) of DO, Bobo-Dioulasso, based on clinical symptoms (fever, headache, chills, abdominal pain, rash, joint pains, muscular pains, vomiting, conjunctival hyperemia, and retroorbital pain). whole blood was collected for each patient into serum separation tubes (3–5 mL), then centrifuged at 1500 rpm for 5 minutes to separate the sera. The serum samples were aliquoted into two separate vials (200 µl/vial) and stored at -80°C before rapid diagnostic test (RDTs), RNA and DNA extraction, molecular detection of DENV, CHIKV, and malaria. RDTs were performed using Dengue Non-Structural 1 (NS1) Antigen and IgG/IgM Antibody (Colloidal Gold, Wondfo) kit and CHIKV IgM/IgG antibodies (Vitrosens Biotechnology RapidFor, Istanbul, Turkey) kit. For each consenting patient, (1–2 mL) of blood samples were collected in ethylene diamine tetra acetic acid (EDTA) tubes and used to screen for malaria infection by a microscopic examination. The protocol of this study was examined and authorized by the Guiriko Burkina Faso Regional Health Department No. 2021 − 0294/MS/RHBS/DRS and the Health Science Research Institute of Burkina Faso ethical committees No. A026-2023/CEIRES/IRSS. All patients who participated in this study gave consent. Molecular screening of DENV1-4 and CHIKV RNA extraction According to the manufacturer's instructions, ribonucleic acid (RNA) was extracted from 140 µL of human serum using the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany) and stored at -80°C until further use. Multiplex RT-PCR for DENV1-4 and CHIKV detection DENV1-4 and CHIKV were screened in RNA samples using the PrimeScript One Step RT-PCR Kit (Takara bio-INC) according to the protocol previously described by Belem et al (2024) [ 31 ]. Briefly, we have prepared a reaction mixture by adding 2,41 µL of H 2 O PCR grade,7.5 µL of 2X RT buffer, 0.45 µL (10 µM) of the conserved forward primer specific to all DENV serotypes (DENV-1, DENV-2, DENV-3, and DENV-4), 0.11 µL (10 µM) of CHIKV forward primer, 0.3 µL (10 µM) of DENV-1 and DENV-4 reverse primers, 0.11 µL(10 µM) of DENV-2, DENV-3, and CHIKV reverse primers, 0.6 µL of the enzyme and 3 µL of RNA. Multiplex RT-PCR was performed in Applied Biosystems (Applied Biosystems™, Thermo Fisher Scientific, Massachusetts, USA). Cycle conditions were 30 min Reverse Transcriptase (RT) at 50°C, followed by 95°C for 5 min, and then 35 cycles of 95°C for 30 s, 62°C for 20 s, 72°C for 30 s, and a final extension of 72°C for 5 min. To interpret PCR results, amplicons were run by gel electrophoresis 1% in 1x Tris-Borate-EDTA buffer with a 100 bp DNA ladder Molecular detection of Plasmodium DNA extraction 50 µL of serum was added to 70 µL of Tris-EDTA 10mM buffer and incubated at 50°C for 30 min on a hot plate (BD ProbeTec TM ET). This was followed by a second incubation at 95°C for 10 min, and centrifugation at 3000 rpm using a Sigma 1–15 machine (Sigma rotor Nr 12124). The supernatant containing DNA was collected in a new 1.5 ml Eppendorf tube and stored at -20°C before molecular detection of Plasmodium Multiplex PCR for Plasmodium species detection Plasmodium was screened using DNA extraction in human serum by polymerase chain reaction (PCR). Plasmodium species were determined using mainly three species circulate in Burkina Faso specific primers (Universal Forward (UF): 5′-GTATCTGATCGTCTTCACTCCC-3′; P. falciparum reverse (Pfr):5’-AACAGACGGGTAGTCATGATTGAG-3’; P. malariae reverse (Pmr): 5’-CGTTAAGATAAACGCCAAGC-3’, and P. ovale reverse (Por):5’ CTGTTCTTTGCATTCCTTATGC-3’). These primers have previously been used in the study of Boonma et al [ 32 ]. The reaction mixture consisted of, 4µL of FIREPol Master Mix (SOLIS BIODYNE), 0.5µL of UF and Pfr primer, 0.6 µL of Pmr and Por primer, 2µL of DNA, and 11.8 µL of H 2 0 PCR grade to complete the final mixture to 20µL. The cycle conditions consist of a 95°C for 5 min, and 35 cycles of 95°C for 30 s, 58°C for 45 s, 72°C for 1 min, and 72°C for 5 min. The PCR products were analyzed by agarose gel electrophoresis 2%. Data management Excel sheet was used to enter clinical, all demographic, and laboratory data until the time of analysis. Each participant was assigned a unique identifier so that names and residential addresses were removed upon entering the questionnaire data into the database, and confidentiality was maintained. Proportions and ratios were calculated for categorical variables such as age, sex, clinical manifestation, molecular detection of the different serotypes of DENV, CHIKV, the different plasmodial species, DENV serology, microscopic examination of malaria, and coinfection. The R version 4.3.1 was used for all analyses. Results Sociodemographic and clinical characteristics A total of 306 patients with the infectious syndrome were included in this study, among whom 52.2% (172/306) were males and 43.8% (134/306) females. The median age was 37 years, ranging from 2 to 84 years (Table 1). The main common symptoms found among patients included in this study were asthenia 95.1% (291/306), headache 92.81% (284/306), fever 88.88% (272/306), muscular pain 73.21% (224 /306), and joint pain 70.51% (216/306) (Table 1). The last reported symptoms were abdominal pain 14.38% (44/306) and retroorbital pain 3.93% (12/306) (Table 1). Rashes and bleeding have not been reported among the patients. Table 1 Clinical and sociodemographic characteristics Variable Sex Number (%) Male 172 (56.2) Female 134 (43.8) Age Years Age median 32 Age range 2 to 84 Clinical symptom Number (%) Headache Yes No 284 (92.81) 22 (7.19) Fever Yes No 272 (88.88) 34 (11.12) Abdominal pain Yes No 44 (14.38) 262 (85.62) Asthenia Yes No 291 (95.1) 15 (4.9) Muscular pain Yes No 224 (73.21) 82 (26.79) Retroorbital pain Yes No 12 (3.93) 294 (96.07) Joint pain Yes No 216 (70.51) 90 (29.41) Rashes Yes No 0 (0.0) 306 (100.0) Bleeding Yes 0 (0.0) No 306 (100.0) Coinfection between probable dengue cases and malaria Out of 306 patients sample analyzed, 4.54% (14/306) were positive for coinfection DENV NS1 and Plasmodium (Microscopy/PCR), 1.96% (6/306) for coinfection DENV (NS1+IgM) and Plasmodium (Microscopy/PCR), 0.98% (3/306) for coinfection DENV (NS1+IgG) and Plasmodium (Microscopy/PCR) (Fig.1). The most coinfection probable dengue cases and malaria are positive DENV IgM and Plasmodium (Microscopy/PCR) with 8.49% (26/306), followed by positive DENV IgG and Plasmodium (Microscopy/PCR) with 7.51 % (23/306) and positive DENV(IgM+IgG) and Plasmodium (Microscopy/PCR) with 5.22% (16/306) (Fig.1). In this study, 71.3% (218/306) of patients were negative for both DENV and Plasmodium or with mono-infection (DENV or Plasmodium ) (Fig.1). Coinfection of acute dengue fever and malaria Among 306 samples screened using DENV RT-PCR and Malaria microscopy detection, 5.22% (16/306) were coinfections (DENV- Plasmodium ) (Fig. 2), 2.94% (9/306) were DENV mono-infections, 45.75% (140/306) were Plasmodium mono-infections, and 46.09% (141/306) were negative for both DENV and Plasmodium . According to DENV screening using RT-PCR and Plasmodium using PCR in samples, 7.51% (23/306) were coinfection (DENV- Plasmodium ) (Fig. 2), 2.94% (9/306) and 44.44% (136/306) were DENV and Plasmodium mono-infection, respectively, and both DENV and Plasmodium negative samples were 45.11% (138/306). In this study, 100% (23/23) of the coinfection sample were Plasmodium -positive by PCR, whereas 69.56 % (16/23) were positive by microscopy. CHIKV has not been detected in this study. Coinfection of DENV serotypes and Plasmodium species In this study, two DENV serotypes (DENV-1 and DENV-3) have been identified (Fig. 3a). The dominant serotype was DENV-3, at 68.0% (17/25), followed by DENV-1 at 32.0% (8/25). DENV-2 and DENV-4 were not identified. P. falciparum and P. malariae are the main species identified in this study (Fig. 3b). P. falciparum 87.3 % (145/166) was the most species, followed by P. malariae 12.7 % (21/166). P. ovale was not identified. Coinfection between DENV-3 and P. falciparum was 74.0% (17/23), 13.0 % (3/23) between DENV-3 and P. malaria, 8.7% (2/23) between DENV-1 and P. falciparum, and 4.3 % (1/23) between DENV-1 and P. malaria (Table 2). Table 2 Coinfection of DENV serotypes and Plasmodium species Species Serotypes DENV-1 % (n/N) DENV-2 % (n/N) DENV-3 % (n/N) DENV-4 % (n/N) P. falciparum 8.7(2/23) 0.0 (0/23) 74.0 (17/23) 0.0 (0/23) P. malaria 4.3 (1/23) 0.0 (0/23) 13.0 (3/23) 0.0 (0/23) P. ovale 0.0 (0/23) 0.0 (0/23) 0.0 (0/23) 0.0 (0/23) Total 13.0 (3/23) 0.0 (0/23) 87.0 (20/23) 0.0 (0/23) Discussion This study occurred in a tropical country (Burkina Faso) during the rainy season, where malaria and arbovirus diseases can spread. This study's main clinical symptoms were asthenia 95.1%, headache 92.81%, fever 88.88%, muscular pain 73.21% and joint pain 70.51%. These symptoms are common to tropical diseases, limiting their clinical diagnosis[ 33 ]. So, clinical identification of arbovirus infections like dengue and malaria coinfection is difficult due to their similar symptoms [ 34 ]. Consequently, this might lead to delayed diagnosis of dengue and malaria co-infections and may cause serious diseases for the patient [ 35 ]. In this study, the proportion of males was higher than females. However, arbovirus and malaria infection risk are not associated with gender-specific but depend on exposure to Aedes and Anopheles mosquito bites [ 36 , 37 ]. However, the outcome of secondary DENV infections is also controlled by sex, with girls > 4 years of age having higher rates of DSS than boys of any age [ 38 ]. This study showed that high coinfection rates were observed in DENV IgM-positive and Plasmodium , at 8.49%, followed by DENV IgG-positive and Plasmodium at 7.51%. Coinfection with DENV NS1-positive and Plasmodium (4.54%) has been found. During the acute phase of infected patients by DENV, the NS1 protein of the virus can be detected in serum samples up to 5–7 days after the symptoms onset, while IgM antibodies are detected from day 5 to 3 months postinfection, and IgG from day 10, till many years later. Usually, anti-DENV IgM antibodies serve as a marker of primary dengue infection, while the secondary infection is detected by an increase of IgG antibodies combined with a lower IgM titer [ 39 ]. Samples positive for DENV IgG probably indicate a previous exposure to any of the DENV serotypes in circulation. The high seroprevalence among the population could also be attributed to past infection, reinfection, ongoing transmission, or increased vector exposure to socioeconomic activities close to mosquito breeding habitats [ 39 ]. Several studies have shown that antibody seropositivity against DENV is notably high in Burkina Faso. These studies have been conducted by Lim et al. (2019) (40.0%), Im et al. (2020) (28.3%) [ 40 , 41 ]. The high dengue seroprevalence and malaria coinfection in participants in this study could reveal the extent of the undiagnosed, misdiagnosed, and hidden burden and prevalence of the two mosquito-borne infections. It could also be the lack of and limited, or underestimated, testing capacities in developing countries health systems and epidemiological serosurveillance facilities [ 42 , 43 ]. In Burkina Faso, many people do not go to the hospital early for medical consultations because of limited financial resources, which may explain the difficulty associated with early detection of DENV in the acute phase The current study has found a 7.51% prevalence of DENV and Plasmodium co-infections among patients by the molecular test of DENV and Plasmodium , against a 5.22% of co-infection DENV molecular test and malaria positive by microscopy. DENV and Plasmodium coinfection are high by molecular tests for both diseases. Light microscopy, the gold standard for laboratory confirmation of malaria, has a sensitivity of detection ranging from 30 to 50 parasites per microliter (p/µL) of blood to 50–500 p/µL [ 44 ]. In addition to having low sensitivity, microscopy depends on the quality of reagents, the techniques used in preparing and staining the smear, and the expertise of the microscopist who examined the smear [ 45 ]. Molecular tools such as polymerase chain reaction (PCR) have higher sensitivity than microscopy and RDT, with a sensitivity of about 2–5 p/µL of blood for Nested PCR and 0.01 to 1 p/µL of blood for real-time PCR [ 46 ]. PCR is very sensitive and can detect Plasmodium species in one reaction using multiplex PCR [ 47 ]. However, DENV and Plasmodium co-infections cause febrile illnesses and present similar symptoms that are not easily differentiated clinically. Therefore, using molecular tools to improve the accuracy of early detection is vital for understanding dengue and malaria co-infections and cocirculation distribution, and for implementing proper therapeutic interventions. The results and information shown by this study can be used as guidance for public health workers, clinicians, and policymakers to choose appropriate strategies, diagnostics, and treatment in Burkina Faso and other endemic areas [ 34 ]. Severe coinfection between dengue and malaria has been reported in several studies [ 35 , 48 , 49 ]. The co-circulation between DENV and Plasmodium involves the coexistence of both disease vectors ( Aedes and Anopheles ) and hosts who may maintain contact with them or migrate to many geographic areas where they are present. Dengue and malaria coinfection can be severe, and fatal consequences can occur; for this reason, it is important to take an adequate and early diagnostic test approach to guarantee their treatment [ 50 ]. A high prevalence of both malaria and dengue coinfections has been reported in Sudan [ 34 ]. Coinfections of dengue and malaria have been reported previously from Cameroon and elsewhere in Africa, at levels similar to the 55.6% were found [ 51 ]. These levels of coinfection with malaria and dengue may reflect the low socio-economic status of the affected population since transmission of both pathogens by their respective vectors is favored by poor housing quality, inadequate sanitation, and poverty [ 52 , 53 ]. In Africa, where malaria transmission rates far exceed anywhere else worldwide, such coinfections will be responsible for substantial underdiagnosis and underreporting of arboviral infections throughout the continent[ 51 ]. Burkina Faso experienced a dengue outbreak in 2023 in a period that was also characterized by a very high number of cases of malaria, which could partly explain the high number of cases of dengue-malaria coinfection and severe dengue cases [ 54 ]. These results suggest that molecular methods and the detection of DENV NS1 by RDT are more useful in the early detection of DENV and Plasmodium in the acute phase, while IgM and IgG serological tests are more indicative of post-viral or past infection of dengue. This highlights the importance of a diagnostic strategy combining molecular and serological methods for detecting dengue at different stages of infection. Burkina Faso Ministry of Health should include molecular diagnostics of dengue and malaria in the health system protocol, mainly in the rainy season, to increase early differential and coinfection diagnostics. In this study, DENV-3 (68.0%) followed by DENV-1 (32.0%) are the main serotypes detected; these serotypes have been responsible for the dengue outbreak in Burkina in 2023 [ 54 ]. For the Plasmodium species, P. falciparum (87.3%) and P. malaria (12.7 %) ave been identified. P. falciparum is the main species involved in malaria cases in Burkina Faso and sub-Saharan Africa [ 55 ]. The most common coinfection in this study was coinfection between DENV-3 and P. falciparum (74.0%). Some studies have shown an association with dengue severity during primary infection and DENV-3 [ 56 , 57 ]. DENV2 and DENV3 caused a higher proportion of severe disease compared to other serotypes [ 58 ]. Increased severity is associated with P. falciparum infection but has also been reported for P. vivax [ 59 ]. The pathogenic mechanisms differ between the two infections; malaria is primarily characterized by anemia due to significant intravascular hemolysis, while thrombocytopenia and fluid leakage are the major features of dengue fever[ 59 ]. Notably, an increase in the incidence of severe thrombocytopenia in patients has been reported during DENV and Plasmodium coinfections [ 59 ]. In this case, both pathologies were characterized as serious due to the clinical manifestations and laboratory findings, which included shock, lung involvement, prostration, myopericarditis, anemia, and thrombocytopenia[ 60 ]. Severe malaria has been reported during coinfection between P. vivax and DENV in Mexico [ 61 ].In 2016, coinfection with DENV-3 and P. vivax was reported in a pediatric patient from New Delhi, India [ 62 ]. Determination of DENV serotype coinfection with plasmodium species is useful in predicting the disease severity. Conclusion Our study demonstrated the usefulness of molecular tools for the early detection of DENV and Plasmodium coinfection. It also showed the co-circulation between DENV-1, DENV-3, P. falciparum , and P. malaria in Bobo-Dioulasso. Early detection of DENV is important for epidemiologic surveillance, predicting outbreaks, and implementing strategies for protecting public health and reducing coinfection of dengue and malaria. This study should be used as guidance for the Burkina Faso Ministry of Health for a new diagnostic algorithm proposal, choosing appropriate strategies, and treatment of dengue and malaria. A new study will be designed to understand more about DENV serotypes and Plasmodium species coinfection in disease severity. Declarations All participants gave consentement and participants with age < 18 years parents gave all consentement. Supplementary Information: Database and CMIC Author Checklist Acknowledgments: We sincerely thank Fond National de la Recherche et de l’Innovation pour le Devéloppement (FONRID) of Burkina Faso, Centre d’Excellence Africain en Innovations Biotechnologiques pour l’Elimination des Maladies à Transmission Vectorielle (CEA/ITECH-MTV) of BURKINA FASO and the World Academy of Science- International Centre for Genetic Engineering and Biotechnology (TWAS-ICGEB). Author contributions Conceptualization: Louis Robert Wendyam Belem, Michel Kiréopori Gomgnimbou, Ibrahim Sangaré. Methodology: Louis Robert Wendyam Belem, Raymond Kharlhis Yao, Miriam Félicité Amara, Armand Vital Wenceslas Taita, Philippe Kaboré, Kouamé Wilfred Ulrich kouadio, Kobo Gnada. Writing – original draft: Louis Robert W. Belem. Writing – review & editing: Sylvester Agha Ibemgbo. Supervision, Project administration: Louis Robert Wendyam Belem, Michel Kiréopori Gomgnimbou, Ibrahim Sangaré Funding This research was supported by the Fond National de la Re­cherche et de l’Innovation pour le Devéloppement of Burkina Faso (Grant No. FONRID/AAP-Spécial-Jeunes/NCP/PCD/2022) and Cen­tre d’Excellence Africain en Innovations Biotechnologiques pour l’Elimination des Maladies à Transmission Vectorielle of Burkina Faso (Grant No.2020 − 000178/MESRSI/SG/UNB/P). Availability of data applicable Conflict of interest The authors declare no conflict of interest. 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Clin Microbiol Infect 25(5):580–585 Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH (2007) A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg 77(6 Suppl):119–127 De Koninck AS, Cnops L, Hofmans M, Jacobs J, Van Den Bossche D, Philippé J (2017) Diagnostic performance of the loop-mediated isothermal amplification (LAMP) based illumigene® malaria assay in a non-endemic region. Malar J 16(1) Belem LRW, Ibemgbo SA, Gomgnimbou MK, Verma DK, Kaboré A, Kumar A et al (2024) Development of Multiplex Molecular Assays for Simultaneous Detection of Dengue Serotypes and Chikungunya Virus for Arbovirus Surveillance. Curr Issues Mol Biol 46(3):2093–2104 Boonma P, Christensen PR, Suwanarusk R, Price RN, Russell B, Lek-Uthai U (2007) Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum. Malar J 6:124 Sow A, Loucoubar C, Diallo D, Faye O, Ndiaye Y, Senghor CS et al (2016) Concurrent malaria and arbovirus infections in Kedougou, southeastern Senegal. Malar J 15(1) Alsedig K, Eldigail MH, Elduma AH, Elaagip A, Altahir O, Siam HA et al (2023) Prevalence of malaria and dengue co-infections among febrile patients during dengue transmission season in Kassala, eastern Sudan. PLoS Negl Trop Dis 17(10):e0011660 Zhao Y, Wu X, Liao F (2018) Severe Cerebral Falciparum Malaria with Dengue Coinfection: A Case Report. Iran J Parasitol 13(2):323–327 Katuwal N, Shrestha A, Ranjitkar U, Jakibanjar S, Kumar S, Tamrakar D et al (2024) Molecular Investigation of DENV serotypes in the dengue outbreak of 2022 in Nepal. Kathmandu Univ Med J 22(85):99–106 Ouédraogo JCRP, Ilboudo S, Compaoré TR, Bado P, Nitiéma M, Ouédraogo WT et al (2024) Determinants and prevalence of symptomatic dengue fever among adults in the Central Region of Burkina Faso: a hospital-based cross-sectional study. BMC Infect Dis 24(1) Guzman MG, Gubler DJ, Izquierdo A, Martinez E, Halstead SB (2016) Dengue infection. Nat Rev Dis Primers 2:16055 Asaga Mac P, Airiohuodion PE, Yako AB, Makpo JK, Kroeger A (2022) The Seroprevalence and Hidden Burden of Chikungunya Endemicity and Malaria Mono- and Coinfection in Nigeria. Int J Environ Res Public Health 19(15) Lim JK, Ridde V, Agnandji ST, Lell B, Yaro S, Yang JS et al (2022) Seroepidemiological Reconstruction of Long-term Chikungunya Virus Circulation in Burkina Faso and Gabon. J Infect Dis 227(2):261–267 Im J, Balasubramanian R, Ouedraogo M, Wandji Nana LR, Mogeni OD, Jeon HJ et al (2020) The epidemiology of dengue outbreaks in 2016 and 2017 in Ouagadougou, Burkina Faso. Heliyon 6(7) Abdullahi IN, Akande AO, Muhammed Y, Rogo LD, Oderinde BS (2020) Prevalence Pattern of Chikungunya Virus Infection in Nigeria: A Four Decade Systematic Review and Meta-analysis. Pathog Glob Health 114(3):111–116 Mala W, Wilairatana P, Kotepui KU, Kotepui M (2021) Prevalence of malaria and chikungunya co-infection in febrile patients: A systematic review and meta-analysis. Trop Med Infect Dis 6(3) Calderaro A, Piccolo G, Chezzi C (2024) The Laboratory Diagnosis of Malaria: A Focus on the Diagnostic Assays in Non-Endemic Areas. Int J Mol Sci 25(2):695 Mathison BA, Pritt BS (2017) Update on malaria diagnostics and test utilization. J Clin Microbiol 55(7):2009–2017 Berzosa P, De Lucio A, Romay-Barja M, Herrador Z, González V, García L et al (2018) Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea. Malar J 17(1) Lazrek Y, Florimond C, Volney B, Discours M, Mosnier E, Houzé S et al (2023) Molecular detection of human Plasmodium species using a multiplex real-time PCR. Sci Rep13(1) Siddig EE, Mohamed NS, Ahmed A (2024) Severe coinfection of dengue and malaria: A case report. Clin Case Rep 12(6) da Cruz MGS, Dos Santos RO, Sousa MGT, Costa FTM, de Lacerda MVG, Lopes SCP et al (2025) Impact of Dengue Virus Infection on the Cytoadherence of Plasmodium vivax-Infected Erythrocytes. Mem Inst Oswaldo Cruz 25:120 González-Macea O, Martínez-Ávila MC, Pérez M, Tibocha Gordon I, Arroyo Salgado B (2023) Concurrent Dengue-Malaria Infection: The Importance of Acute Febrile Illness in Endemic Zones. Clin Med Insights Case Rep 16:11795476221144585 Tchetgna HS, Yousseu FS, Kamgang B, Tedjou A, McCall PJ, Wondji CS (2021) Concurrent circulation of dengue serotype 1, 2 and 3 among acute febrile patients in cameroon. PLoS Negl Trop Dis 15(10) Piedrahita LD, Agudelo Salas IY, Marin K, Trujillo AI, Osorio JE, Arboleda-Sanchez SO et al (2018) Risk Factors Associated with Dengue Transmission and Spatial Distribution of High Seroprevalence in Schoolchildren from the Urban Area of Medellin, Colombia. Can J Infect Dis Med Microbiol 2018:2308095 Araújo DdaC, Dos Santos AD, Lima SVMA, Vaez AC, Cunha JO, de Araújo KCGM (2020) Determining the association between dengue and social inequality factors in north-eastern Brazil: A spatial modelling. Geospat Health 15(1) Gomgnimbou MK, Belem LRW, Some K, Diallo M, Barro B, Kaboré A et al (2024) Utilization of novel molecular multiplex methods for the detection and, epidemiological surveillance of dengue virus serotypes and chikungunya virus in Burkina Faso, West Africa. Mol Biol Rep 51(1) Bonko Mdit, Tahita A, Kiemde MC, Lompo F, Mens P, Tinto PF H, et al (2022) Diagnostic Performance of Plasmodium falciparum Histidine-Rich Protein-2 Antigen-Specific Rapid Diagnostic Test in Children at the Peripheral Health Care Level in Nanoro (Burkina Faso). Trop Med Infect Dis 7(12) Tatura SNN, Denis D, Santoso MS, Hayati RF, Kepel BJ, Yohan B et al (2021) Outbreak of severe dengue associated with DENV-3 in the city of Manado, North Sulawesi, Indonesia. Int J Infect Dis 106:185–196 Hernández Bautista PF, Cabrera Gaytán DA, Santacruz Tinoco CE, Vallejos Parás A, Alvarado Yaah JE, Martínez Miguel B et al (2024) Retrospective Analysis of Severe Dengue by Dengue Virus Serotypes in a Population with Social Security, Mexico 2023. Viruses 16(5) Narvaez F, Montenegro C, Juarez JG, Zambrana JV, Gonzalez K, Videa E et al (2025) Dengue severity by serotype and immune status in 19 years of pediatric clinical studies in Nicaragua. PLoS Negl Trop Dis 19(1):e0012811 Mendonça VRR, Andrade BB, Souza LCL, Magalhães BML, Mourão MPG, Lacerda MVG et al (2015) Unravelling the patterns of host immune responses in Plasmodium vivax malaria and dengue co-infection. Malar J 14(1) Queb-Pech NM, Núñez-Oreza LA, Estrada-Méndez A, Tamay-Segovia P, Collí-Heredia JP, Blum-Domínguez SC (2022) Unusual co-infection of severe malaria by Plasmodium vivax and dengue virus in Mexico. Trop Biomed 39(4):575–578 Soo KM, Khalid B, Ching SM, Chee HY (2016) Meta-analysis of dengue severity during infection by different dengue virus serotypes in primary and secondary infections. PLoS ONE 11(5) Tazeen A, Abdullah M, Hisamuddin M, Ali S, Naqvi IH, Verma HN et al (2017) Concurrent Infection with Plasmodium vivax and the Dengue and Chikungunya Viruses in a Paediatric Patient from New Delhi, India in 2016. Intervirology 60(1–2):48–52 Additional Declarations No competing interests reported. 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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-7658090","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":520086758,"identity":"afd4d82e-93be-43d2-87a7-cdf22df6a081","order_by":0,"name":"Louis Robert Wendyam 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Ulrich","lastName":"kouadio","suffix":""},{"id":520086764,"identity":"2f844483-6170-407b-8f57-398d2a867cff","order_by":6,"name":"Kobo Gnada","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Kobo","middleName":"","lastName":"Gnada","suffix":""},{"id":520086765,"identity":"219f9073-2dcf-43b8-9fe4-f1aa7b016aba","order_by":7,"name":"Sylvester Agha Ibemgbo","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Sylvester","middleName":"Agha","lastName":"Ibemgbo","suffix":""},{"id":520086766,"identity":"839d7f2d-5ded-4b8c-9492-536fbf142c8d","order_by":8,"name":"Ibrahim Sangaré","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Ibrahim","middleName":"","lastName":"Sangaré","suffix":""},{"id":520086767,"identity":"182c1873-c578-40d7-8e14-4377fb229275","order_by":9,"name":"Michel Kiréopori Gomgnimbou","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Michel","middleName":"Kiréopori","lastName":"Gomgnimbou","suffix":""}],"badges":[],"createdAt":"2025-09-19 11:23:33","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7658090/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7658090/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":92478812,"identity":"51e961f7-1204-4848-82d3-c1359f255285","added_by":"auto","created_at":"2025-09-30 07:35:25","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":23872,"visible":true,"origin":"","legend":"\u003cp\u003eProbable dengue cases and \u003cem\u003ePlasmodium \u003c/em\u003ecoinfection\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7658090/v1/a60299e2a15f7be1168b2c0a.jpg"},{"id":92478813,"identity":"abcc4125-2785-44aa-91ac-2d17eec84aab","added_by":"auto","created_at":"2025-09-30 07:35:25","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":29609,"visible":true,"origin":"","legend":"\u003cp\u003eCoinfection of DENV and \u003cem\u003ePlasmodium\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7658090/v1/b10883e698526dac50765a71.jpg"},{"id":92478814,"identity":"41f6d49b-5f2b-43d4-b93f-7581cf022da6","added_by":"auto","created_at":"2025-09-30 07:35:25","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":25245,"visible":true,"origin":"","legend":"\u003cp\u003eMolecular detection of DENV and \u003cem\u003ePlasmodium. \u003c/em\u003e\u003cstrong\u003ea \u003c/strong\u003eDENV serotypes\u003cstrong\u003e \u003c/strong\u003ein circulation. \u003cstrong\u003eb \u003c/strong\u003eThe two main \u003cem\u003ePlasmodium\u003c/em\u003especies detected in this study. M (100 bp DNA marker); DENV-1 (200 bp); DEN-3 (1359 bp); \u003cem\u003eP. falciparum \u003c/em\u003e(276 bp); \u003cem\u003eP. malariae \u003c/em\u003e(411 bp); bp: base pair; PC: positive control; NC: negative control.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7658090/v1/5eb51d21168885cbee078140.jpg"},{"id":92480776,"identity":"586ebdd5-4c38-4bd6-986b-e17a33c1dbca","added_by":"auto","created_at":"2025-09-30 07:43:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":911874,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7658090/v1/5d92371d-9c32-4827-9990-c7bd7b3e0da4.pdf"},{"id":92478815,"identity":"578acb8b-f0f9-4266-bec2-b5efa2f3e8a0","added_by":"auto","created_at":"2025-09-30 07:35:25","extension":"xlsx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":45536,"visible":true,"origin":"","legend":"","description":"","filename":"CoinfectiondatabaseBELEM.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7658090/v1/5fabe0013ce1f2821d386101.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Importance of molecular tools in arbovirus and malaria disease coinfection detection in humans, Bobo- Dioulasso, western Burkina Faso","fulltext":[{"header":"Introduction","content":"\u003cp\u003eArbovirus infection and malaria are vector-borne diseases and are responsible for major public health issues in the tropical and subtropical regions, including Burkina Faso [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Dengue virus (DENV) and chikungunya virus (CHIKV) are the main arboviruses in the world and are transmitted mainly by \u003cem\u003eAedes (Ae) aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. More than 3.9\u0026nbsp;billion humans in the world are susceptible to contracting DENV, principally in tropical countries [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. According to the World Health Organization (WHO), CHIKV is now present in more than 110 countries across Africa, Asia, and the Americas [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. CHIKV and DENV are ribonucleic acid (RNA) viruses belonging to the \u003cem\u003eTogaviridae\u003c/em\u003e and \u003cem\u003eFlaviviridae\u003c/em\u003e families, respectively. DENV is classified into four genetically different serotypes (DENV1-4) [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Prior infection by one DENV serotype offers long-term immunity to that serotype. However, subsequent infection by a different serotype can cause severe disease like dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Generally, CHIKV infection has low mortality rates. However, it can cause significant morbidity, impacting the quality of life for infected individuals and causing economic losses, mainly in the least developed countries [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/p\u003e\u003cp\u003eMalaria is a parasite disease, mainly caused by \u003cem\u003ePlasmodium (P.) falciparum\u003c/em\u003e, \u003cem\u003eP. vivax\u003c/em\u003e, \u003cem\u003eP. malariae\u003c/em\u003e, and \u003cem\u003eP. ovale\u003c/em\u003e. The \u003cem\u003ePlasmodium\u003c/em\u003e genome is Deoxyribonucleic (DNA) [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. In sub-Saharan Africa, \u003cem\u003eP. falciparum\u003c/em\u003e is the most common parasite responsible for 90% of the global malaria burden and all malaria deaths[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The \u003cem\u003eAnopheles\u003c/em\u003e mosquito transmits malaria, and \u003cem\u003eAnopheles gambiae\u003c/em\u003e is considered to be the essential malaria transmission vector in Africa [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. It is expected that the highly mobile lifestyle of the population, the increased activities made possible by dependable international transportation networks, and climatic change will increase the prevalence of dengue and malaria co-infection [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMalaria and arbovirus infections presented similar main symptoms, such as fever, headache, myalgia, arthralgia, rash, nausea, diarrhea, vomiting, and abdominal pain [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Malaria is endemic and the first disease responsible for mortality in Burkina Faso. In Burkina, the DENV is the main arbovirus responsible for outbreaks with high prevalence and important morbidity and mortality. Between August and November 2023, Burkina Faso reported important DENV outbreaks and fewer CHIKV outbreaks [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eSeveral studies have shown that DENV and Malaria can co-circulate [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Consequently, the co-circulation of both diseases complicates the diagnosis and treatment process [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. In Burkina Faso, many people use antimalarial treatment for a fever or headache without medical consultation and biological diagnosis. This practice can lead to disease complications if the patient is infected by DENV or coinfected with malaria. Although malaria or DENV mono infections can be sometimes severe, coinfections between both diseases could be even more fatal [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Because of their similar clinical manifestations, probable concurrent dengue fever and malaria are often neglected and could lead to misdiagnosis as malaria alone [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. During coinfection, misdiagnosis is probably higher than mono-infection; consequently, slow identification of dengue outbreaks can occur and lead to high morbidity or mortality.\u003c/p\u003e\u003cp\u003eSerological diagnosis of arbovirus is limited because antibodies such as immunoglobulin M (IgM) and immunoglobulin G (IgG) may not be detectable early in the acute phase of the disease, and cross-reactivity between anti-flavivirus antibodies may occur [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Many commercial rapid diagnostic tests (RDTs) to detect CHIKV and DENV are available [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. However, false-positive test results have been reported in some studies, which is a limitation in the diagnosis [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Several studies have reported that sometimes malaria is misdiagnosed by rapid diagnostic tests (RDTs) due to Histidine-Rich Protein 2 (HRP2) and HRP3 gene deletions [\u003cspan additionalcitationids=\"CR27\" citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Microscopic examination of Giemsa-stained blood slides has been the standard for malaria diagnosis for nearly a century. However, this technique is labor-intensive, time-consuming, and challenged by a high limit of detection (LoD). Under optimal circumstances, the latter is predicted to be nearly 50 parasites/\u0026micro;L and is highly dependent on the slide's quality and the microscopist's level of expertise [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Furthermore, personnel\u0026rsquo;s lack of practice and proficiency may account for delays and errors in diagnosis. However, molecular methods are the best means of differential diagnosis and can detect coinfection between vector-borne diseases[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. In Burkina, DENV and \u003cem\u003ePlasmodium\u003c/em\u003e coinfection data are scarce in healthcare and less described due to the diagnostic algorithm shortcoming. This study was designed to demonstrate the importance of molecular tools in the early detection of coinfection, as well as the co-circulation of arbovirus infection, and malaria in Bobo-Dioulasso. Early detection of coinfection between arbovirus and malaria is crucial for appropriate treatment and essential to prevent severe disease and death.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eSite and study design\u003c/h2\u003e\u003cp\u003eThis is a cross-sectional study, performed in Bobo-Dioulasso (11\u0026deg; 11\u0026prime; 00\u0026Prime; North, 4\u0026deg; 17\u0026prime; 00\u0026Prime; West), belonging to the Guiriko region, Burkina Faso, during the rainy season between June 2023 and August 2023. The study period offers good conditions for mosquito breeding, such as \u003cem\u003eAnopheles\u003c/em\u003e and \u003cem\u003eAedes\u003c/em\u003e, that is favorable for malaria and arbovirus transmission. Participants have been included in the medical centers with surgical antenna (CMA) of DO, Bobo-Dioulasso, based on clinical symptoms (fever, headache, chills, abdominal pain, rash, joint pains, muscular pains, vomiting, conjunctival hyperemia, and retroorbital pain). whole blood was collected for each patient into serum separation tubes (3\u0026ndash;5 mL), then centrifuged at 1500 rpm for 5 minutes to separate the sera. The serum samples were aliquoted into two separate vials (200 \u0026micro;l/vial) and stored at -80\u0026deg;C before rapid diagnostic test (RDTs), RNA and DNA extraction, molecular detection of DENV, CHIKV, and malaria. RDTs were performed using Dengue Non-Structural 1 (NS1) Antigen and IgG/IgM Antibody (Colloidal Gold, Wondfo) kit and CHIKV IgM/IgG antibodies (Vitrosens Biotechnology RapidFor, Istanbul, Turkey) kit. For each consenting patient, (1\u0026ndash;2 mL) of blood samples were collected in ethylene diamine tetra acetic acid (EDTA) tubes and used to screen for malaria infection by a microscopic examination. The protocol of this study was examined and authorized by the Guiriko Burkina Faso Regional Health Department No. 2021\u0026thinsp;\u0026minus;\u0026thinsp;0294/MS/RHBS/DRS and the Health Science Research Institute of Burkina Faso ethical committees No. A026-2023/CEIRES/IRSS. All patients who participated in this study gave consent.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eMolecular screening of DENV1-4 and CHIKV\u003c/h3\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003eRNA extraction\u003c/h2\u003e\u003cp\u003eAccording to the manufacturer's instructions, ribonucleic acid (RNA) was extracted from 140 \u0026micro;L of human serum using the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany) and stored at -80\u0026deg;C until further use.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eMultiplex RT-PCR for DENV1-4 and CHIKV detection\u003c/h3\u003e\n\u003cp\u003eDENV1-4 and CHIKV were screened in RNA samples using the PrimeScript One Step RT-PCR Kit (Takara bio-INC) according to the protocol previously described by Belem et \u003cem\u003eal\u003c/em\u003e (2024) [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Briefly, we have prepared a reaction mixture by adding 2,41 \u0026micro;L of H\u003csub\u003e2\u003c/sub\u003eO PCR grade,7.5 \u0026micro;L of 2X RT buffer, 0.45 \u0026micro;L (10 \u0026micro;M) of the conserved forward primer specific to all DENV serotypes (DENV-1, DENV-2, DENV-3, and DENV-4), 0.11 \u0026micro;L (10 \u0026micro;M) of CHIKV forward primer, 0.3 \u0026micro;L (10 \u0026micro;M) of DENV-1 and DENV-4 reverse primers, 0.11 \u0026micro;L(10 \u0026micro;M) of DENV-2, DENV-3, and CHIKV reverse primers, 0.6 \u0026micro;L of the enzyme and 3 \u0026micro;L of RNA. Multiplex RT-PCR was performed in Applied Biosystems (Applied Biosystems\u0026trade;, Thermo Fisher Scientific, Massachusetts, USA). Cycle conditions were 30 min Reverse Transcriptase (RT) at 50\u0026deg;C, followed by 95\u0026deg;C for 5 min, and then 35 cycles of 95\u0026deg;C for 30 s, 62\u0026deg;C for 20 s, 72\u0026deg;C for 30 s, and a final extension of 72\u0026deg;C for 5 min. To interpret PCR results, amplicons were run by gel electrophoresis 1% in 1x Tris-Borate-EDTA buffer with a 100 bp DNA ladder\u003c/p\u003e\u003cp\u003e\u003cb\u003eMolecular detection of\u003c/b\u003e \u003cb\u003ePlasmodium\u003c/b\u003e\u003c/p\u003e\n\u003ch3\u003eDNA extraction\u003c/h3\u003e\n\u003cp\u003e50 \u0026micro;L of serum was added to 70 \u0026micro;L of Tris-EDTA 10mM buffer and incubated at 50\u0026deg;C for 30 min on a hot plate (BD ProbeTec \u003csup\u003eTM\u003c/sup\u003e ET). This was followed by a second incubation at 95\u0026deg;C for 10 min, and centrifugation at 3000 rpm using a Sigma 1\u0026ndash;15 machine (Sigma rotor Nr 12124). The supernatant containing DNA was collected in a new 1.5 ml Eppendorf tube and stored at -20\u0026deg;C before molecular detection of \u003cem\u003ePlasmodium\u003c/em\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eMultiplex PCR for\u003c/b\u003e \u003cb\u003ePlasmodium\u003c/b\u003e \u003cb\u003especies detection\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003ePlasmodium\u003c/em\u003e was screened using DNA extraction in human serum by polymerase chain reaction (PCR). Plasmodium species were determined using mainly three species circulate in Burkina Faso specific primers (Universal Forward (UF): 5\u0026prime;-GTATCTGATCGTCTTCACTCCC-3\u0026prime;; \u003cem\u003eP. falciparum\u003c/em\u003e reverse (Pfr):5\u0026rsquo;-AACAGACGGGTAGTCATGATTGAG-3\u0026rsquo;; \u003cem\u003eP. malariae\u003c/em\u003e reverse (Pmr): 5\u0026rsquo;-CGTTAAGATAAACGCCAAGC-3\u0026rsquo;, and \u003cem\u003eP. ovale\u003c/em\u003e reverse (Por):5\u0026rsquo; CTGTTCTTTGCATTCCTTATGC-3\u0026rsquo;). These primers have previously been used in the study of Boonma et \u003cem\u003eal\u003c/em\u003e [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The reaction mixture consisted of, 4\u0026micro;L of FIREPol Master Mix (SOLIS BIODYNE), 0.5\u0026micro;L of UF and Pfr primer, 0.6 \u0026micro;L of Pmr and Por primer, 2\u0026micro;L of DNA, and 11.8 \u0026micro;L of H\u003csub\u003e2\u003c/sub\u003e0 PCR grade to complete the final mixture to 20\u0026micro;L. The cycle conditions consist of a 95\u0026deg;C for 5 min, and 35 cycles of 95\u0026deg;C for 30 s, 58\u0026deg;C for 45 s, 72\u0026deg;C for 1 min, and 72\u0026deg;C for 5 min. The PCR products were analyzed by agarose gel electrophoresis 2%.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eData management\u003c/h2\u003e\u003cp\u003eExcel sheet was used to enter clinical, all demographic, and laboratory data until the time of analysis. Each participant was assigned a unique identifier so that names and residential addresses were removed upon entering the questionnaire data into the database, and confidentiality was maintained. Proportions and ratios were calculated for categorical variables such as age, sex, clinical manifestation, molecular detection of the different serotypes of DENV, CHIKV, the different plasmodial species, DENV serology, microscopic examination of malaria, and coinfection. The R version 4.3.1 was used for all analyses.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eSociodemographic and clinical characteristics\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of 306 patients with the infectious syndrome were included in this study, among whom 52.2% (172/306) were males and 43.8% (134/306) females. The median age was 37 years, ranging from 2 to 84 years (Table 1). The main common symptoms found among patients included in this study were asthenia 95.1% (291/306), headache 92.81% (284/306), fever 88.88% (272/306), muscular pain 73.21% (224 /306), and joint pain 70.51% (216/306) (Table 1). The last reported symptoms were abdominal pain 14.38% (44/306) and retroorbital pain 3.93% (12/306) (Table 1). Rashes and bleeding have not been reported among the patients.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Clinical and sociodemographic characteristics\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVariable\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSex\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003eNumber (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eMale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e172 (56.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eFemale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e134 (43.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003eYears\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eAge median\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eAge range\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e2 to 84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eClinical symptom\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003eNumber (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eHeadache\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;284 (92.81)\u003c/p\u003e\n \u003cp\u003e22 (7.19)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eFever\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e272 (88.88)\u003c/p\u003e\n \u003cp\u003e34 (11.12)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eAbdominal pain\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e44 (14.38)\u003c/p\u003e\n \u003cp\u003e262 (85.62)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eAsthenia\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e291 (95.1)\u003c/p\u003e\n \u003cp\u003e15 (4.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eMuscular pain\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e224 (73.21)\u003c/p\u003e\n \u003cp\u003e82 (26.79)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eRetroorbital pain\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e12 (3.93)\u003c/p\u003e\n \u003cp\u003e294 (96.07)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eJoint pain\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e216 (70.51)\u003c/p\u003e\n \u003cp\u003e90 (29.41)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eRashes\u003c/p\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003e0 (0.0)\u003c/p\u003e\n \u003cp\u003e306 (100.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eBleeding\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e0 (0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56.279%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43.2559%;\"\u003e\n \u003cp\u003e306 (100.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eCoinfection between probable dengue cases and malaria\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOut of 306 patients sample analyzed, 4.54% (14/306) were positive for coinfection DENV NS1 and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR), 1.96% (6/306) for coinfection DENV (NS1+IgM) and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR), 0.98% (3/306) for coinfection DENV (NS1+IgG) and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR) (Fig.1). The most coinfection probable dengue cases and malaria are positive DENV IgM and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR) with 8.49% (26/306), followed by positive DENV IgG and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR) with 7.51 % (23/306) and positive DENV(IgM+IgG) and \u003cem\u003ePlasmodium\u003c/em\u003e (Microscopy/PCR) with 5.22% (16/306) (Fig.1). In this study, 71.3% (218/306) of patients were negative for both DENV and \u003cem\u003ePlasmodium\u003c/em\u003e or with mono-infection (DENV or \u003cem\u003ePlasmodium\u003c/em\u003e) (Fig.1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCoinfection of acute dengue fever and malaria\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAmong 306 samples screened using DENV RT-PCR and Malaria microscopy detection, 5.22% (16/306) were coinfections (DENV-\u003cem\u003ePlasmodium\u003c/em\u003e) (Fig. 2), 2.94% (9/306) were DENV mono-infections, 45.75% (140/306) were \u003cem\u003ePlasmodium\u003c/em\u003e mono-infections, and 46.09% (141/306) were negative for both DENV and \u003cem\u003ePlasmodium\u003c/em\u003e. According to DENV screening using RT-PCR and \u003cem\u003ePlasmodium\u003c/em\u003e using PCR in samples, 7.51% (23/306) were coinfection (DENV-\u003cem\u003ePlasmodium\u003c/em\u003e) (Fig. 2), 2.94% (9/306) and 44.44% (136/306) were DENV and \u003cem\u003ePlasmodium\u003c/em\u003e mono-infection, respectively, and both DENV and \u003cem\u003ePlasmodium\u003c/em\u003e negative samples were 45.11% (138/306). In this study, 100% (23/23) of the coinfection sample were \u003cem\u003ePlasmodium\u003c/em\u003e-positive by PCR, whereas 69.56 % (16/23) were positive by microscopy. CHIKV has not been detected in this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCoinfection of DENV serotypes and \u003cem\u003ePlasmodium\u003c/em\u003e species\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn this study, two DENV serotypes (DENV-1 and DENV-3) have been identified (Fig. 3a). The dominant serotype was DENV-3, at 68.0% (17/25), followed by DENV-1 at 32.0% (8/25). DENV-2 and DENV-4 were not identified. \u003cem\u003eP. falciparum\u003c/em\u003e and \u003cem\u003eP. malariae\u0026nbsp;\u003c/em\u003eare the main species identified in this study (Fig. 3b). \u003cem\u003eP. falciparum\u003c/em\u003e 87.3 % (145/166) was the most species, followed by \u003cem\u003eP. malariae\u0026nbsp;\u003c/em\u003e12.7 % (21/166). \u003cem\u003eP. ovale\u0026nbsp;\u003c/em\u003ewas not identified.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCoinfection between DENV-3 and \u003cem\u003eP. falciparum\u0026nbsp;\u003c/em\u003ewas 74.0% (17/23), 13.0 % (3/23) between DENV-3 and \u003cem\u003eP. malaria,\u0026nbsp;\u003c/em\u003e8.7% (2/23) between DENV-1 and \u003cem\u003eP. falciparum,\u0026nbsp;\u003c/em\u003eand 4.3\u003cem\u003e\u0026nbsp;%\u003c/em\u003e (1/23) between DENV-1 and \u003cem\u003eP. malaria\u0026nbsp;\u003c/em\u003e(Table 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e Coinfection of DENV serotypes and \u003cem\u003ePlasmodium\u003c/em\u003e species\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"637\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerotypes\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDENV-1\u0026nbsp;\u003c/strong\u003e% (n/N)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDENV-2\u0026nbsp;\u003c/strong\u003e% (n/N)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDENV-3\u0026nbsp;\u003c/strong\u003e% (n/N)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDENV-4\u0026nbsp;\u003c/strong\u003e% (n/N)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eP. falciparum\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e8.7(2/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e74.0 (17/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eP. malaria\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.3 (1/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e13.0 (3/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eP. ovale\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e13.0 (3/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e87.0 (20/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e0.0 (0/23)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study occurred in a tropical country (Burkina Faso) during the rainy season, where malaria and arbovirus diseases can spread. This study's main clinical symptoms were asthenia 95.1%, headache 92.81%, fever 88.88%, muscular pain 73.21% and joint pain 70.51%. These symptoms are common to tropical diseases, limiting their clinical diagnosis[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. So, clinical identification of arbovirus infections like dengue and malaria coinfection is difficult due to their similar symptoms [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Consequently, this might lead to delayed diagnosis of dengue and malaria co-infections and may cause serious diseases for the patient [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. In this study, the proportion of males was higher than females. However, arbovirus and malaria infection risk are not associated with gender-specific but depend on exposure to \u003cem\u003eAedes\u003c/em\u003e and \u003cem\u003eAnopheles\u003c/em\u003e mosquito bites [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. However, the outcome of secondary DENV infections is also controlled by sex, with girls\u0026thinsp;\u0026gt;\u0026thinsp;4 years of age having higher rates of DSS than boys of any age [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThis study showed that high coinfection rates were observed in DENV IgM-positive and \u003cem\u003ePlasmodium\u003c/em\u003e, at 8.49%, followed by DENV IgG-positive and \u003cem\u003ePlasmodium\u003c/em\u003e at 7.51%. Coinfection with DENV NS1-positive and \u003cem\u003ePlasmodium\u003c/em\u003e (4.54%) has been found. During the acute phase of infected patients by DENV, the NS1 protein of the virus can be detected in serum samples up to 5\u0026ndash;7 days after the symptoms onset, while IgM antibodies are detected from day 5 to 3 months postinfection, and IgG from day 10, till many years later. Usually, anti-DENV IgM antibodies serve as a marker of primary dengue infection, while the secondary infection is detected by an increase of IgG antibodies combined with a lower IgM titer [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Samples positive for DENV IgG probably indicate a previous exposure to any of the DENV serotypes in circulation. The high seroprevalence among the population could also be attributed to past infection, reinfection, ongoing transmission, or increased vector exposure to socioeconomic activities close to mosquito breeding habitats [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Several studies have shown that antibody seropositivity against DENV is notably high in Burkina Faso. These studies have been conducted by Lim et al. (2019) (40.0%), Im et al. (2020) (28.3%) [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. The high dengue seroprevalence and malaria coinfection in participants in this study could reveal the extent of the undiagnosed, misdiagnosed, and hidden burden and prevalence of the two mosquito-borne infections. It could also be the lack of and limited, or underestimated, testing capacities in developing countries health systems and epidemiological serosurveillance facilities [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. In Burkina Faso, many people do not go to the hospital early for medical consultations because of limited financial resources, which may explain the difficulty associated with early detection of DENV in the acute phase\u003c/p\u003e\u003cp\u003eThe current study has found a 7.51% prevalence of DENV and \u003cem\u003ePlasmodium\u003c/em\u003e co-infections among patients by the molecular test of DENV and \u003cem\u003ePlasmodium\u003c/em\u003e, against a 5.22% of co-infection DENV molecular test and malaria positive by microscopy. DENV and \u003cem\u003ePlasmodium\u003c/em\u003e coinfection are high by molecular tests for both diseases. Light microscopy, the gold standard for laboratory confirmation of malaria, has a sensitivity of detection ranging from 30 to 50 parasites per microliter (p/\u0026micro;L) of blood to 50\u0026ndash;500 p/\u0026micro;L [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. In addition to having low sensitivity, microscopy depends on the quality of reagents, the techniques used in preparing and staining the smear, and the expertise of the microscopist who examined the smear [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Molecular tools such as polymerase chain reaction (PCR) have higher sensitivity than microscopy and RDT, with a sensitivity of about 2\u0026ndash;5 p/\u0026micro;L of blood for Nested PCR and 0.01 to 1 p/\u0026micro;L of blood for real-time PCR [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. PCR is very sensitive and can detect \u003cem\u003ePlasmodium\u003c/em\u003e species in one reaction using multiplex PCR [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. However, DENV and \u003cem\u003ePlasmodium\u003c/em\u003e co-infections cause febrile illnesses and present similar symptoms that are not easily differentiated clinically. Therefore, using molecular tools to improve the accuracy of early detection is vital for understanding dengue and malaria co-infections and cocirculation distribution, and for implementing proper therapeutic interventions. The results and information shown by this study can be used as guidance for public health workers, clinicians, and policymakers to choose appropriate strategies, diagnostics, and treatment in Burkina Faso and other endemic areas [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Severe coinfection between dengue and malaria has been reported in several studies [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. The co-circulation between DENV and \u003cem\u003ePlasmodium\u003c/em\u003e involves the coexistence of both disease vectors (\u003cem\u003eAedes\u003c/em\u003e and \u003cem\u003eAnopheles\u003c/em\u003e) and hosts who may maintain contact with them or migrate to many geographic areas where they are present. Dengue and malaria coinfection can be severe, and fatal consequences can occur; for this reason, it is important to take an adequate and early diagnostic test approach to guarantee their treatment [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. A high prevalence of both malaria and dengue coinfections has been reported in Sudan [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Coinfections of dengue and malaria have been reported previously from Cameroon and elsewhere in Africa, at levels similar to the 55.6% were found [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. These levels of coinfection with malaria and dengue may reflect the low socio-economic status of the affected population since transmission of both pathogens by their respective vectors is favored by poor housing quality, inadequate sanitation, and poverty [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. In Africa, where malaria transmission rates far exceed anywhere else worldwide, such coinfections will be responsible for substantial underdiagnosis and underreporting of arboviral infections throughout the continent[\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Burkina Faso experienced a dengue outbreak in 2023 in a period that was also characterized by a very high number of cases of malaria, which could partly explain the high number of cases of dengue-malaria coinfection and severe dengue cases [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThese results suggest that molecular methods and the detection of DENV NS1 by RDT are more useful in the early detection of DENV and \u003cem\u003ePlasmodium\u003c/em\u003e in the acute phase, while IgM and IgG serological tests are more indicative of post-viral or past infection of dengue. This highlights the importance of a diagnostic strategy combining molecular and serological methods for detecting dengue at different stages of infection. Burkina Faso Ministry of Health should include molecular diagnostics of dengue and malaria in the health system protocol, mainly in the rainy season, to increase early differential and coinfection diagnostics.\u003c/p\u003e\u003cp\u003eIn this study, DENV-3 (68.0%) followed by DENV-1 (32.0%) are the main serotypes detected; these serotypes have been responsible for the dengue outbreak in Burkina in 2023 [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. For the \u003cem\u003ePlasmodium\u003c/em\u003e species, \u003cem\u003eP. falciparum\u003c/em\u003e (87.3%) and \u003cem\u003eP. malaria\u003c/em\u003e (12.7 %) ave been identified. \u003cem\u003eP. falciparum\u003c/em\u003e is the main species involved in malaria cases in Burkina Faso and sub-Saharan Africa [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]. The most common coinfection in this study was coinfection between DENV-3 and \u003cem\u003eP. falciparum\u003c/em\u003e (74.0%). Some studies have shown an association with dengue severity during primary infection and DENV-3 [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. DENV2 and DENV3 caused a higher proportion of severe disease compared to other serotypes [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Increased severity is associated with \u003cem\u003eP. falciparum\u003c/em\u003e infection but has also been reported for \u003cem\u003eP. vivax\u003c/em\u003e [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. The pathogenic mechanisms differ between the two infections; malaria is primarily characterized by anemia due to significant intravascular hemolysis, while thrombocytopenia and fluid leakage are the major features of dengue fever[\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. Notably, an increase in the incidence of severe thrombocytopenia in patients has been reported during DENV and \u003cem\u003ePlasmodium\u003c/em\u003e coinfections [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. In this case, both pathologies were characterized as serious due to the clinical manifestations and laboratory findings, which included shock, lung involvement, prostration, myopericarditis, anemia, and thrombocytopenia[\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. Severe malaria has been reported during coinfection between \u003cem\u003eP. vivax\u003c/em\u003e and DENV in Mexico [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e].In 2016, coinfection with DENV-3 and \u003cem\u003eP. vivax\u003c/em\u003e was reported in a pediatric patient from New Delhi, India [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. Determination of DENV serotype coinfection with \u003cem\u003eplasmodium\u003c/em\u003e species is useful in predicting the disease severity.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eOur study demonstrated the usefulness of molecular tools for the early detection of DENV and \u003cem\u003ePlasmodium\u003c/em\u003e coinfection. It also showed the co-circulation between DENV-1, DENV-3, \u003cem\u003eP. falciparum\u003c/em\u003e, and \u003cem\u003eP. malaria\u003c/em\u003e in Bobo-Dioulasso. Early detection of DENV is important for epidemiologic surveillance, predicting outbreaks, and implementing strategies for protecting public health and reducing coinfection of dengue and malaria. This study should be used as guidance for the Burkina Faso Ministry of Health for a new diagnostic algorithm proposal, choosing appropriate strategies, and treatment of dengue and malaria. A new study will be designed to understand more about DENV serotypes and \u003cem\u003ePlasmodium\u003c/em\u003e species coinfection in disease severity.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAll participants gave consentement and participants with age \u0026lt; 18 years parents gave all consentement.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eSupplementary Information:\u0026nbsp;\u003c/strong\u003eDatabase and\u0026nbsp;CMIC Author Checklist\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003eWe sincerely thank Fond National de la Recherche et de l\u0026rsquo;Innovation pour le Dev\u0026eacute;loppement (FONRID) of Burkina Faso, Centre d\u0026rsquo;Excellence Africain en Innovations Biotechnologiques pour l\u0026rsquo;Elimination des Maladies \u0026agrave; Transmission Vectorielle (CEA/ITECH-MTV) of BURKINA FASO and the World Academy of Science- International Centre for Genetic Engineering and Biotechnology (TWAS-ICGEB).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003eConceptualization: Louis Robert Wendyam Belem, Michel Kir\u0026eacute;opori Gomgnimbou, Ibrahim Sangar\u0026eacute;. Methodology:\u0026nbsp;Louis Robert Wendyam Belem, Raymond Kharlhis Yao, Miriam F\u0026eacute;licit\u0026eacute; Amara, Armand Vital Wenceslas Taita, Philippe Kabor\u0026eacute;, Kouam\u0026eacute; Wilfred Ulrich kouadio, Kobo Gnada. Writing \u0026ndash; original draft: Louis Robert W. Belem. Writing \u0026ndash; review \u0026amp; editing: Sylvester Agha Ibemgbo. Supervision, Project administration: Louis Robert Wendyam Belem, Michel Kir\u0026eacute;opori Gomgnimbou, Ibrahim Sangar\u0026eacute;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e This research was supported by the Fond National de la Re\u0026shy;cherche et de l\u0026rsquo;Innovation pour le Dev\u0026eacute;loppement of Burkina Faso (Grant No. FONRID/AAP-Sp\u0026eacute;cial-Jeunes/NCP/PCD/2022) and Cen\u0026shy;tre d\u0026rsquo;Excellence Africain en Innovations Biotechnologiques pour l\u0026rsquo;Elimination des Maladies \u0026agrave; Transmission Vectorielle of Burkina Faso (Grant No.2020 \u0026minus; 000178/MESRSI/SG/UNB/P).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data\u0026nbsp;\u003c/strong\u003eapplicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e The authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e: The protocol of this study was first reviewed and approved by the institutional ethics committee of Health Science Research, Burkina Faso, No. A026-2023/CEIRES/IRSS. All human serum samples were obtained with patients\u0026apos; informed consent.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eShah PD, Mehta TK (2017) Evaluation of concurrent malaria and dengue infections among febrile patients. Indian J Med Microbiol 35(3):402\u0026ndash;405\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu W, Wang J, Yu N, Yan J, Zhuo Z, Chen M et al (2018) Development of multiplex real-time reverse-transcriptase polymerase chain reaction assay for simultaneous detection of Zika, dengue, yellow fever, and chikungunya viruses in a single tube. J Med Virol 90(11):1681\u0026ndash;1686\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSekaran SD, Artsob H (2007) Molecular diagnostics for the detection of human flavivirus infections. 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Intervirology 60(1\u0026ndash;2):48\u0026ndash;52\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Molecular arbovirus malaria coinfection Bobo-Dioulasso","lastPublishedDoi":"10.21203/rs.3.rs-7658090/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7658090/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn tropical regions, arbovirus disease and malaria co-circulate currently; consequently, co-infection of both diseases can be found and complicates the diagnosis and treatment process with potentially high morbidity and mortality. This study was designed to demonstrate the co-circulation of arbovirus infection and malaria in Bobo-Dioulasso, as well as the importance of molecular tools in the early detection of coinfection. This cross-sectional study was conducted in Bobo-Dioulasso, Burkina Faso, between June 2023 and August 2023. Participants were included based on clinical symptoms, and blood samples were collected for dengue rapid diagnostic test (RDTs), molecular detection of dengue virus, chikungunya virus, and malaria. Microscopic examination was also performed to diagnose malaria infection. Among 306 samples screened using DENV RT-PCR and Malaria microscopy detection, 5.22% (16/306) were DENV-\u003cem\u003ePlasmodium\u003c/em\u003e coinfections. According to DENV screening using RT-PCR and malaria screening using PCR, 7.51% (23/306) were found to be coinfected with DENV and \u003cem\u003ePlasmodium\u003c/em\u003e. In this study, 100% (23/23) of the coinfection samples were malaria-positive by PCR, whereas 69.56% (16/23) were positive by microscopy. CHIKV has not been detected in this study. Among coinfections, 74.0% (17/23) were coinfections between DENV-3 and \u003cem\u003eP. falciparum\u003c/em\u003e, 13.0% (3/23) between DENV-3 and \u003cem\u003eP. malariae\u003c/em\u003e, 8.7% (2/23) between DENV-1 and \u003cem\u003eP. falciparum\u003c/em\u003e, and 4.3% (1/23) between DENV-1 and \u003cem\u003eP. malariae.\u003c/em\u003e Our study demonstrated the utility of molecular tools in detecting dengue and malaria coinfection in the acute phase. It also showed the co-circulation between DENV-1, DENV-3, \u003cem\u003eP. falciparum, and P. malariae.\u003c/em\u003e\u003c/p\u003e","manuscriptTitle":"Importance of molecular tools in arbovirus and malaria disease coinfection detection in humans, Bobo- Dioulasso, western Burkina Faso","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-30 07:35:21","doi":"10.21203/rs.3.rs-7658090/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"32cb2444-224b-4dfb-aa3c-79f80eaf4439","owner":[],"postedDate":"September 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-30T07:35:21+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-30 07:35:21","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7658090","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7658090","identity":"rs-7658090","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}