A critical appraisal on antibody detector-serodiagnostics for Bovine Tuberculosis

A critical appraisal on antibody detector-serodiagnostics for Bovine Tuberculosis | 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 A critical appraisal on antibody detector-serodiagnostics for Bovine Tuberculosis Rafik Soliman, Neven Waheeb, Essam Nasr, Mahmoud El-hariri, Heidy Abo-Elyazeed, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2121571/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 Bovine tuberculosis is a worldwide disease that causes great economic losses to the dairy industry and constitutes a serious human public health hazard. In the present study, two types of commercially available immunochromatographic lateral flow kits, namely, IQRT Anigen Rapid Bovine TB Ab kit and Ubio quick VET Bovine Tuberculosis Antibody kit were evaluated for their sero-diagnostic sensitivity and specificity of bovine tuberculosis through the detection of Mycobacterium bovis -specific antibodies in tested serum samples. The results obtained by these kits were compared with the ELISA test, bacteriological examination, and tuberculin skin testing. In this study, out of 3750 dairy cattle tested by single intradermal tuberculin skin test, 69 (1.8%) animals reacted positively. From the slaughtered tuberculin-positive cattle 51 animals (73.9%) showed visible lesions on postmortem examination while the remaining 18 animals (26.1%) showed no visible lesions. Using bacteriological examination of tissue specimens collected, 48 out of the 69 positive tuberculin reactor animals (69.6%) were positive and M. bovis was successfully isolated. Using ELISA for serological examination of serum samples collected prior to skin testing from the 69 positive tuberculin reactors, only 26 cases (37.7%) were positive. The serodiagnosis of bovine tuberculosis using the IQRT Anigen lateral flow kit, which employed recombinant M. bovis MPB70 antigen as capture and detector, could identify only 21 positive cases, which represent 43.8% of 48 bacteriologically identified cases and 30.44% out of the 69 tuberculin positive cases. On the other hand, the Ubio quick VET kit has detected zero% of bovine tuberculosis-positive cattle. It is concluded that the use bovine tuberculosis Antibody Rapid Test Kit alone may be unreliable for the detection of tuberculosis-infected cattle. Bovine tuberculosis tuberculin skin test M. bovis isolation lateral flow kits for antibody detection ELISA Rapid diagnostics Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Bovine tuberculosis (bTB) is a chronic granulomatous disease mainly affecting lymph nodes and lung tissues of cattle. It is caused by M. bovis , a member of M. tuberculosis complex [ 1 ]. M. bovis zoonotically transmitted to humans; causing disease through ingestion; inhalation and less commonly by contact with mucous membranes and broken skin [ 2 ]. Bovine tuberculosis is characterized by the progressive development of granulomatous lesions (tubercles) in any body organ and can affect a broadly large number of species [ 3 ]. The intradermal tuberculin skin test has been in use for almost a century in the diagnosis of bovine tuberculosis and despite the technological advances of the last two decades, it is still the only reliable test for the diagnosis of tuberculosis in cattle [ 4 ]. The Purified Protein Derivative (PPD), the antigen used in the tuberculin test, contains many antigenic determinants of broad specificity, which leads to the appearance of non-specific reactors indicating false positive possibilities [ 5 ]. Although the tuberculin test is still considered the golden master test for diagnosis of bovine tuberculosis, it is tedious, laborious and, time-consuming. In addition, negative or doubtful tuberculin-tested cases cannot be retested before 6 months after tuberculin testing. Therefore, the recently developed serodiagnostic methods may be suitable and reasonable alternative tools for the diagnosis of bovine tuberculosis. Therefore, the suggestion of [ 6 ] to use of ELISA with the tuberculin skin test to overcome the problems of the tuberculin test was very convincing. The first ELISA kit for the sero-diagnosis of bovine tuberculosis was introduced by [ 7 ], and it was for wildlife application using PPD-B as the target antigen. Previously developed ELISA kit was reported to have a sensitivity and specificity of 73% and 96%, respectively, for the detection of TB in wild boar [ 8 ]. As an alternative to tuberculin skin testing for antemortem diagnosis, quick and simple immune-chromatographic LFA assays for the serodiagnosis of bTB have been developed and presented. [ 9 , 10 ]. As both qualitative captures and detectors of specific antibodies against M. bovis in plasma, serum, and whole blood, these chromatographic immunoassays use special blends of chosen M. bovis antigens. [ 11 , 12 ]. The common sero-reactive antigens in bovine TB have been identified as the MPB83, 14-kDa protein, ESAT-6, MPB70, CFP-10, MPT51, MPT32, MPB59, MPB64, MPT63, Acr1, PstS-1, 16-kDa alpha-crystalline/MPB83 fusion protein, ESAT-6/CFP10 fusion protein, M. bovis purified protein derivatives, and M. bovis culture filtrate [ 11 , 13 ]. The use of LFK in the sero-diagnosis of infectious diseases has been widely applied and its first application in the diagnosis of bovine tuberculosis was first recorded by [ 14 ]. Since then, these tests have become an extremely popular tool for the rapid diagnosis of bovine tuberculosis [ 8 ]. However, the specificity and sensitivity of several commercially available LFA kits are questionable. In the present work, two LFKs were evaluated in the rapid detection of tuberculosis-infected cattle. The sensitivity and specificity of these kits were determined using the tuberculin skin test and bacterial isolation as a golden standard technique. The first kit was the IQRT Antigen Rapid Bovine TB antibody kit that employed recombinant M. bovis MPB70 antigen (specific for M. bovis ) as capture and detector. The second kit was the Ubio quick VET Bovine TB Antibody Rapid Test, this kit uses a unique cocktail of recombinant antigens of M. bovis to detect specific antibodies of three immunoglobulin classes, IgG, IgM, and IgA developed during infection in cattle. Material And Methods Study design and animal population Through a national study involving most of the Egyptian governorates, a total of 3750 crossbreed dairy cattle from different Egyptian Governorates, all over Egypt were involved in the current study. The study design can be clearly described through the flowchart illustrated in Figure (1) . Application of the single intradermal Tuberculin skin test done according to the OIE manual (2009) as follows; Testing of each animal started with measuring skin thickness followed by an I/D injection 0.1 ml of bovine PPD. Seventy-two hours post-injection, the skin thickness was measured and the differences between the 1 st and 2 nd reading in (mm) was recorded. According to the Egyptian General Organization of Veterinary Services [15] an increase in the skin thickness of 4mm or more was considered positive, less than 2mm is considered as negative and from 3- 4 mm was considered as doubtful. All positive reactants were slaughtered due to the “test and slaughter rule” in case of TB positive feedlot animal cases. Isolation of acid-fast bacilli from collected tissue samples of slaughtered tuberculin positive reactants: The tested tuberculin-positive cattle were slaughtered. Postmortem examination was conducted on them, and 69 tissue samples were collected. These samples were used for bacteriological isolation and identification of M. bovis according to [16] . Organs, lymph nodes, and/or other tissues were processed for the isolation of this organism. The collected sample processing was done as follows; all specimens were cut into small pieces, mixed with fine sterile sand, diluted with 2ml sterile distilled water, and ground in a sterile mortar. Two ml of 4% H 2 SO 4 were added to the samples, which were then incubated for ½ hour at 37 o C. The mixture was furtherly diluted with 16 ml sterile distilled water and centrifuged at 3000 rpm/20 minutes. The obtained sediment was resuspended in 0.5 ml sterile distilled water, inoculated into Lowenstein-Jensen slants, and incubated at 37 o C in an inclined position overnight and then vertically for at least 6-8 weeks with weekly examination starting from three days post inoculation and the obtained results were recorded [17]. Serological diagnosis of bovine tuberculosis using locally developed ELISA and 2 types of commercially available Lateral flow Kits: All sero-diagnostic tests were performed on the serum samples that were collected from the examined dairy cattle before being tested with the single intradermal tuberculin skin test [18] . These samples were examined for diagnosis of bovine tuberculosis through the detection Mycobacterium bovis -specific antibodies. The serum samples were examined using a locally prepared ELISA and two commercially available lateral flow kits, namely; the Antigen Rapid Bovine TB Ab test Kit (IQRT test kits) and the Ubio quick VET Bovine Tuberculosis Antibody Rapid Test Kit . The obtained results were comparably analyzed. - Examination of the collected serum samples using the Anigen Rapid Bovine TB Ab test Kit (IQRT test kits) produced by Anigen Animal Genetics Inc., South Korea: The test was performed per the manufacturer’s instructions as follows; three drops (about 60 - 90μl) of tested serum samples were dispensed into the sample application well. One drop of phosphate buffered saline was added into the sample well if the flow migration is not observed within 30 seconds in the result window, Results was read in 20 minutes with instructional invalid readings and interpretation after 20 minutes. Positive results appeared as two purple color bands (T band and C band) within the result window, positive results can be visible in as short as 1 minute. If the intensity of the purple band color is faint, it is interpreted as positive as it appears within 20 minutes. - Examination of the collected serum samples using the Ubio quick VET Bovine Tuberculosis Antibody Rapid Test Kit produced by Ubio, India: The test was performed per the manufacturer’s instructions as follows; three drops (about 60 - 90μl) of the tested serum were added to the sample hole on the test card using a dropper. The result was recorded in 5-10 minutes. The control line, marked C, should always develop color if the test procedure is performed properly and the test reagents are working. A red test line marked T will be visible if there are enough Bovine TB specific antibodies in the serum sample. - Examination of the collected serum samples using locally prepared ELISA: The antigen used for coating the microtiter plate wells was bovine PPD (B-PPD) that was obtained from the Bacteriological Diagnostic Products Department, Veterinary Serum and Vaccine Research Institute (VSVRI), Abbasia, Cairo, Egypt. It was prepared according to the protocol of the Central Veterinary Laboratories, Weighbridge, United Kingdom. Briefly, the test was developed as follows; Polystyrene 96-well microtiter plates were coated with the B-PPD antigen using 15µg/ml in 0.1M carbonate-bicarbonate buffer (pH 9.6). The antigen solution was dispensed in 50ul/well. In each microtiter plate 4 wells were left uncoated with B-PPD antigen and served as blank control wells. Each of these blank wells received 50µl of carbodiimide in 0.1 M carbonate bicarbonate coating buffer. The plates were incubated overnight at 4°C then decanted, washed 3 times with PBS and air dried for 10 minutes. Then all wells except the blank ones were filled 100µl/well of 0.01 M ammonium chloride as blocking buffer and further incubated for 30 minutes at 22ºC. Finally, the plates were washed 3 times with PBS washing buffer [19] . In each microtiter plate, all wells except the wells of the first column were filled with 50ul/well PBS buffer. For each serum sample one row is used. The tested serum samples were diluted 1:40 in PBS and 100µl of each tested serum sample was added to the first well of the corresponding row. This was followed by two-fold serial dilutions of each sample. Each plate received negative serum samples run in the same dilutions to calculate the cut-off value. The plates were incubated at room temperature for 30 minutes on a horizontal shaker. The microtiter plates were decanted, washed 4 times with ELISA washing buffer and allowed to stand inverted for 30minutes. To each well, 50µl of anti-bovine IgG alkaline phosphatase diluted 1:5000 in diluting buffer were added and the plates were then incubated for 30 minutes at room temperature on a horizontal shaker. The plates were again washed 4 times with ELISA washing buffer and allowed to stand inverted for 30 minutes. The substrate solution composed of P- Nitrate phenyl phosphate (one tablet 5mg /5ml substrate buffer) was prepared and 50µl/well was dispensed in all wells of the plates and were further incubated for 20min at room temperature. The color development was stopped by the addition of 100µl/well of 0.4 M NaOH-stopping solution. The results were recorded by optical density measurement at 405 nm using spectra III ELISA reader. A titer of 1:80 or higher was considered positive for bovine tuberculosis [20]. The results obtained by the applied serological tests were compared with the tuberculin testing and the results of the culture isolation, which is considered the gold standard test. The diagnostic value of the tested serological methods was determined in the terms of accuracy index, sensitivity, specificity, positive predictive value, and negative predictive value. Results And Discussion Bovine tuberculosis (bTB) is a zoonotic disease with public health significance. The maximum detection of bovine TB in cattle populations in Egypt is vital to understand its epidemiology and zoonotic potentials and to achieve significant reduction and control of the disease in livestock. The tuberculin skin tests (TST) are currently the best available techniques for international field diagnosis of bTB in live animals [2, 21] and it is based on delayed hypersensitivity reactions. The results of tuberculin skin testing are presented in In Table (1) where out of 3750 tuberculin-tested cross-bred dairy cattle, 69 were found to be positive reactors with a prevalence rate of 1.8%. Farm Total no. of examined animals Positive tuberculin Number % 1 170 2 1.2 2 220 5 2.3 3 290 7 2.4 4 140 3 2.1 5 320 8 2.5 6 500 12 2.4 7 410 11 2.7 8 250 3 1.2 9 180 2 1.1 10 290 4 1.4 11 370 5 1.4 12 155 2 1.3 13 225 2 0.9 14 110 2 1.8 15 120 1 0.83 Total 3750 69 1.8 Table (1): Tuberculin skin test result in dairy farm cattle from some Egyptian Governorates. This is comparatively lower than that recorded by other investigators in Egypt [22], (6.9%), [23], ( 4.6%), and [24], (2.2%) and in some other African countries as in Ethiopia (11.6%) [25], and in Chad (8%) [26] . This noticable reduced incidence might be attributed to the fact that the farms included in this research perform the tuberculin test regularly and applied test and slaughter strategy [27] . On the other hand, the prevalence rate recorded in the present study is comparatively higher than that reported by other investigators in other countries, 0.9% to 1.3% in Tanzania [28, 29], respectively, 0.54 % in Venezuela [30] , 0.4 %, in New Zealand [31] 0.05 to 0.15 % in Australi a [32] and 0.02 % in Japan [33] . The postmortem findings of the slaughtered TB reactors revealed that 73.9% of these animals showed visible lesions. At the same time the percentage of tuberculin reactors with non-visible lesions were 26.1%, Table (2) . Total No. of animals Visible lesions Non- visible lésions Generalized Head Pulmonary Digestive Mixed 69 No. % No. % No. % No. % No. % No. % 8 11.6 5 7.2 22 31.9 6 8.7 10 14.5 18 26.1 Table (2): Result of Postmortem examination of tuberculin positive slaughtered cattle. These results agreed with those reported by [34, 35] . On the other hand, a much higher percentage was recorded by [36] who reported (86.9%) in Queensland, [37]. In Egypt Lower percentage was also reported by [38], (46%), [39], (52.7%), and [40], (40.8%). The percentage of tuberculin positive animals with non-visible lesions was attributed to sensitization by atypical mycobacteria or even closely related microorganisms especially the members of the genus Nocardia or a combination of Liver Fluke infestation with saprophytic mycobacteria [41, 42]. Moreover, [43, 44] attributed the cause of non-specific reaction to the assumption that, those animals might be slaughtered at early stage of the disease where the tuberculous lesions were still invisible, or lesions may be found in parts of the body not routinely examined in the carcass such as bone or brain. The relation between the postmortem findings in different ages of tuberculin reactor cattle is presented in Table (3). Table (3): Results of correlation between age of animal and PM finding of the slaughtered cases The percentage of reaction-positive animals increased with age, reaching a maximum in animals over 60 months old. Similar studies in Great Britain recorded an increase of incidence of bTB with increased age [45] . It was suggested earlier by [46] that, the high positivity of the old age might be due to the reduced number γ and δ T cells responsible of the anti-mycobacterial immunity. [These cells are predominantly found in the circulation of young calves. It has also been suggested that increased incidence of TB in older animals can be due to a waning of protective capability in aged animals [47, 48] . The correlation between site of lesion and the isolated mycobacteria species is shown in Table (4). No. of processed tissue samples Bacteriological isolation (52 isolates) M. bovis Unidentified mycobacteria No. % Slow growers Rapid growers No. % No. % 69 48 69.6 3 4.3 1 1.4 Table (4): Result of bacteriological isolation and identification of mycobacteria isolated from tuberculin positive cattle. The total acid-fast bacilli isolated from the 69 slaughtered cattle were 52 isolates (75.4%). According to the morphological characters, growth rate, pigmentation, growth at different temperatures and biochemical tests these isolates were identified into 48 (69.6%) M. bovis , 1 unidentified rapid grower strain (1.44%) and 3 (4.3%) unidentified pigmented slow grower strains. The recovery rate of M. bovis figured up to 69.6% but other authors reported a lower M. bovis recovery rates, 14.8% by [49], 20.2% by [50], 41% by [51 ], 35.4% by [52], 29.1% by [53] , 5.6% by [54] , and 42.9% by [55] . On the contrary, [56] , in Korea reported a much higher isolation rate (92.1%), in the republic of Korea. These results depend mainly on the actual disease status present in the tested herd and to some extent on the experience of the investigators as well as the technique used for decontamination of tissue specimens. Data presented in Figure (2) , shows the relation between the post-slaughter findings of positive tuberculin reactors and the type of mycobacteria recovered from the 69 carcasses, M. bovis isolates , rapid growers isolates and slow growers pigmented isolates were recovered at rates of 69.6 %, 4.3%, and 1.44 %, respectively. M. bovis was isolated at a rate of 100% from generalized TB, 91% from pulmonary TB, 81.8% from Extra Pulmonary TB , 90% from mixed TB, and 11.1% from non-visible lesion (NVL) reactors. These results coincide with those reported by [57] who isolated M. bovis from NVL reactors. On the other hand, the recovery rate of typical mycobacteria rapid growers and slow growers pigmented was 16.7% of the extra-pulmonary for both of them and 33.3% of the mixed for the rapid growers. None of the cases of generalized tuberculosis yielded atypical mycobacteria, since M. bovis has a tendency to induce progressive lesions and generalization [44, 58] . All cases with generalized TB yielded M. bovis, a finding indicating the high susceptibility of cattle to M. bovis infection. Comparison between bacteriological examination and serological testing by lateral flow kits of tuberculin-positive cattle are shown in Figure (3) . The results of the application of lateral flow kits on sera from tuberculin-positive cattle : The One- step Anigen Rapid Bovine Antibody Test (IQRT) employed recombinant M. bovis MPB70 antigen specific for M. bovis as capture and detector antigen. It is important to also mention that serological testing was conducted prior to TST because the latter can boost antibody responses in infected cattle and emphasizes the importance of timing of collection of blood samples on the interpretation the test [59]. This Anigen TB antibody test kit has detected 43.8% of tuberculin positive cattle against 69.6% that were confirmed by bacterial isolation of M. bovis . While the other lateral flow assay Ubio quick VET test is a single-directional lateral-flow serological test, which was produced to provide a quick determination of the presence of M. bovis antibody [60, 61] has detected zero % of tuberculin positive cattle. Comparison between the result of ELISA test, rapid lateral flow test and bacteriological isolation of tuberculin positive cattle showed that 8 tuberculin positive reactors with generalized tuberculous lesions were bacteriologically positive (M. bovis isolation) and ELSIA positive using bovine PPD as coating antigen (100%). As shown in Figure (4) , out of 22 tuberculin positive reactors 10 (45.5%) showed pulmonary lesion, out of 11 tuberculin positive reactors 3 (27.3%) showed extra-pulmonary lesions (head and digestive), and Out of 10 tuberculin reactors showing mixed TB lesions (pulmonary and extra pulmonary), 5 (50%) isolates of M. bovis were positive for ELISA by using bovine PPD as coating antigens. Out of 18 tuberculin reactors showing NVL, 2 (11.1%) isolates of M. bovis could be isolated from them and zero (0%) serum samples were positive by ELISA by using bovine PPD as coating antigen. The results obtained in the current study agreed with those reported by [62, 63] who stated that, antibody response to M. bovis infection is certainly not uniform. This phenomenon was previously suggested in serological analysis by [64] who studied the humoral immune response to M. bovis infection in cattle and concluded that, it was characterized by highly heterogeneous antigen recognition. It is concluded that single intradermal tuberculin test, culture and isolation of the mycobacterium are recommended for determination of a true prevalence of bTB in the herd to be established. The recent lateral flow rapid kits could be used for initial tuberculosis screening in combination with tuberculin skin test (TST) for improving sensitivity of bovine tuberculosis screening, thereby leading to more successful control programs in developing countries. The rapid test is proposed as a potentially useful ancillary assay for bTB. In addition, it may be most suitable for surveillance, especially if an immediate result is needed. The differences in the sensitivity and specificity of the different LFA kits sources available in the market necessitate that, the importance of choosing type of diagnostic kits used for rapid detection of bovine tuberculosis. Also, these kits alone may be not enough for accurate diagnosis of bovine tuberculosis. Declarations Ethics approval and consent to participate The current conducted study is reported in accordance with (Animal Research: Reporting of In-Vivo Experiments-ARRIVE) guidelines. The guidelines of the (Institutional Animal Care and Use Committee-IACUC of faculty of veterinary medicine, Cairo University) were completely followed during any procedures involving animal use through the current conducted study. This study is approved by the ethical committee of the institutional animal care and use committee (IACUC) of Veterinary-Cairo University. No anesthesia or euthanasia protocols were used with the animal involved during this study as all animal-dependent methodological procedures were considered as no to low pain-causing procedures that ethically can be done on a conscious alive animal. All sample collection procedures were done based on permission from the cattle farm owners and managers. Consent for publication 'Not applicable' Availability of data and materials The datasets used and/or analyzed related to the animal cases tested during the current study are available from the corresponding author upon reasonable request. Competing interests The authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article. Funding 'Not applicable' Authors' contributions All authors contributed equally during the whole stages of the study. Acknowledgments We thank all general practitioner veterinarians and dairy animal specialists all over Egypt due to their help during the sample collection stages. References Taylor GM, Worth DR, Palmer S, Jahans K, Hewinson RG. (2007): Rapid detection of mycobacterium bovis DNA in cattle lymph nodes with visible lesions using PCR.BMC Vet Res. 2007 Jun 13;3: 12. De la Rua-Domenech R. 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Analysis of the postmortem finding in tuberculin positive cattle slaughtered at Lodz abattoir during the period 1966–1969. Edycyna Wet. 1970;26:545. Yoon YD, Kim JM, Son BH. Detection of bovine tuberculosis and the factors affecting tuberculin reaction. Korean J Vet Public Heal. 1979;3(1):1–8. Rodriguez VD, Real VF, Gasca AA. Bovine tuberculosis in Cordoba Province of Spain. Hygia Pecovis. 1983;5:31–7. Waddington FG. Observation on tuberculin sensitivity in cattle in Kenya. Br Vet J. 1965;121:319–31. Andrew W, Byrne J, Graham C, Brown A, Donaghy M, Guelbenzu-Gonzalo J, McNair R, Skuce A, Allen, Stanley McDowell (2017): Bovine tuberculosis visible lesions in cattle culled during herd breakdowns: the effects of individual characteristics, trade movement and co-infection. BMC Veterinary Research volume 13(400) DOI 10.1186/s12917-017-1321-z . O'Reilly LM. (1992): [[Specificity and sensitivity of tuberculin tests. Pr Int. Conf. Anim. Tuberculosis, General Organization of Vet. Ser., Cairo, Egypt, P.87–139. Huitema H. Tuberculosis in animals and man 1 st Ed. Publication of the Royal Netherlands Tuberculosis Association (KNCV); 1994. Pollock Ellen Brooks-. Conlan AJK, Mitchell AP, Blackwell R, McKinley TJ, James LN Wood (2013): Age-dependent patterns of bovine tuberculosis in cattle http://www.veterinaryresearch.org/content/44/1/97 . Mackay CR, Hein WR. A large proportion of bovine T cells express the T cell receptor and show a distinct tissue distribution and surface phenotype. Int Immunol. 1989;1:540–5. O'Reilly LM, Daborn CJ. The epidemiology of Mycobacterium bovis infections in animals and man: a review. Tuberc Lung Dis. 1995;76:1–146. Parlas M, Rossi L. Causes of recurrence of tuberculosis in TB free herds. Vet Med. 1964;9:1–10. Lesslie IW, Birn KJ. Mycobacterium aviam infections in cattle and pigs in Great Britain. Tuber Land. 1970;51:446–51. Beck G, Bibrack B. Pathological, histological, and cultural diagnosis of mycobacterial infections with special reference to meat inspection regulations. Tierarztl Umschau. 1971;26(5):196–201. Osman K. (1974): Studies on acid fast micro-organisms in some domesticated animals with special reference to atypical mycobacteria group. Ph. D. (Microbiology) thesis, Cairo University. Gallo JAG, Veitia F, Remon S, Delgado L. (1983): Mycobacteria isolated from cattle reacted to tuberculin tests. Revista-Cubana de CIENCIAS Vet., 14 (3): pp. 173–6. Payeur JB, Marquardt JG. (1988): Mycobacterial isolated from submissions at NVSL National Veterinary Services Laboratories during Fy 83–87. Proceeding of the United States Animal Health Association 92: 457–477. Abuo-Eisha AM, El-Attar AA, El-Sheary MN. Bovine and atypical mycobacterial infection of cattle and buffalos in Port Said Province, Egypt. Assiut Vet. Med J. 1995;47:152–62. Chul Soon Choi. Mycobacterium isolated from man, animals, and soil in Korea, 1968–1980 review. Korean J Vet Public Heal. 1981;5(1):49–64. Rogers RJ, Donad BA, Suchlt K. The distribution of M. bovis in Queens and cattle herds with observation on the laboratory diagnosis of tuberculosis. Assuit Vet J. 1980;56(11):542–6. Good M, Bakker D, Duignan A, Collins DM. The History of In Vivo Tuberculin Testing in Bovines: Tuberculosis, a “One Health” Issue. Front Vet Sci. 2018;5:59. doi: 10.3389/fvets.2018.00059 . Palmer MV, Waters WR, Thacker TC, Greenwald R, Esfandiari J, Lyashchenko KP. Effects of different tuberculin skin-testing regimens on gamma interferon and antibody responses in cattle experimentally infected with Mycobacterium bovis. Clin Vaccine Immunol. 2006;13(3):387–94. ” ( . Harrington NP, Surujballi OP, Prescott JF, Duncan JR, Waters WR, Lyashchenko K, Greenwald R. Antibody responses of cervids (Cervus elaphus) following experimental Mycobacterium bovis infection and the implications for immunodiagnosis. Clin Vaccine Immunol. 2008;15:1650–8. Danbrini S, Sackey AKB, Ayo JO, Bawa EK, Kudi AC, Okaiyeto SO, Pewan SB. Exposer and shedding in milk of Mycobacterium bovis in dairy herds using one-step Anigen rapid bovine tuberculosis antibodies test and Ziehl-Neelsen stain. Medwell J Vet Res. 2010;3(3):38–42. Danbirni S, Okaiyeto SO, Bature C, Moris A. Field Determination of Tuberculosis Prevalence in a Herd of Cattle Using Tuberculin and Quicking® Bovine Tuberculosis Antibody Rapid Tests in Jalingo. Nigeria J Vet Adv. 2013;3(1):20–3. Lepper AWD, corner LA. (1983): Naturally Occurring Mycobacteriosis of Animals in Biology of Mycobacteria. Vol 2 Academic Pres London. P. 478–488. – 546. Waters WR, Whelan AO 2, Lyashchenko KP, Greenwald R 3, Palmer MV, Harris BN, Hewinson RG, Vordermeier HM. (2010): immune responses in cattle inoculated with mycobacterium bovis, mycobacterium tuberculosis, or mycobacterium kansasii. clinical and vaccine immunology, feb. 2010, p. 247–252. 1556–6811 /10/ $ 12.00 doi:10.1128/cvi.00442-09. Hanna J, Neill SD, O’Brien JJ. ELISA tests for antibodies in experimental bovine tuberculosis. Vet Microbiol. 1992;31:243–9. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-2121571","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":143378599,"identity":"a973f3ba-b47b-4378-8f36-928cdffc0bac","order_by":0,"name":"Rafik Soliman","email":"data:image/png;base64,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","orcid":"","institution":"Cairo University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Rafik","middleName":"","lastName":"Soliman","suffix":""},{"id":143378600,"identity":"0d4452b0-2c4d-447d-953d-1d333851add6","order_by":1,"name":"Neven Waheeb","email":"","orcid":"","institution":"Vacsera (Egypt)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Neven","middleName":"","lastName":"Waheeb","suffix":""},{"id":143378601,"identity":"28af5d2c-a576-4093-9629-d9e1eb8a8a6c","order_by":2,"name":"Essam Nasr","email":"","orcid":"","institution":"Vacsera (Egypt)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Essam","middleName":"","lastName":"Nasr","suffix":""},{"id":143378602,"identity":"92984b76-026e-4440-aa88-6c928a3b94e8","order_by":3,"name":"Mahmoud El-hariri","email":"","orcid":"","institution":"Cairo University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mahmoud","middleName":"","lastName":"El-hariri","suffix":""},{"id":143378603,"identity":"ce062e84-f7f1-43ef-9b7b-03583f897971","order_by":4,"name":"Heidy Abo-Elyazeed","email":"","orcid":"","institution":"Cairo University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Heidy","middleName":"","lastName":"Abo-Elyazeed","suffix":""},{"id":143378604,"identity":"7d9bffcd-6daa-4847-8f87-e7df4cc57d2a","order_by":5,"name":"Hassan Aboul-Ella","email":"","orcid":"","institution":"Cairo University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hassan","middleName":"","lastName":"Aboul-Ella","suffix":""}],"badges":[],"createdAt":"2022-09-30 18:30:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2121571/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2121571/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":27789136,"identity":"6810f2b9-1b43-4f8c-83e5-91aeb7f6ba94","added_by":"auto","created_at":"2022-10-14 17:39:53","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":150972,"visible":true,"origin":"","legend":"\u003cp\u003eA flowchart illustrating the overall testing flow of each animal involved in the study\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2121571/v1/549095b14267f4828c96ea91.jpeg"},{"id":27789134,"identity":"db4d1962-35f7-49c2-bf89-f4d3915444a8","added_by":"auto","created_at":"2022-10-14 17:39:53","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":5761,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation between site of lesion and the isolated \u003cem\u003emycobacteria spp.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-2121571/v1/53194b740e97ecdaa9717cec.png"},{"id":27789137,"identity":"0337d78d-1618-4596-848a-d496f4bfa473","added_by":"auto","created_at":"2022-10-14 17:39:53","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":45750,"visible":true,"origin":"","legend":"\u003cp\u003eComparison between bacteriological examination and serological testing (lateral flow kits) of tuberculin positive cattle.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-2121571/v1/0c567c96cd837e96100a11c5.png"},{"id":27789135,"identity":"9ba5f193-e3ea-42bc-8e54-ef8eb04f1b1a","added_by":"auto","created_at":"2022-10-14 17:39:53","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":6749,"visible":true,"origin":"","legend":"\u003cp\u003eComparison between the result of ELISA test, rapid lateral flow test and bacteriological isolation of tuberculin positive cattle. MOTT; Mycobacteria Other Than Tuberculosis.\u003c/p\u003e","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-2121571/v1/a3d92a9effd82654de4df568.png"},{"id":29128583,"identity":"c6144319-e4ae-4c4d-ae98-2ff5015e1b2d","added_by":"auto","created_at":"2022-11-16 10:29:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":788559,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2121571/v1/819f2b7e-8aac-45cd-a7e5-8badb04b2078.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"A critical appraisal on antibody detector-serodiagnostics for Bovine Tuberculosis","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eBovine tuberculosis (bTB) is a chronic granulomatous disease mainly affecting lymph nodes and lung tissues of cattle. It is caused by \u003cem\u003eM. bovis\u003c/em\u003e, a member of \u003cem\u003eM. tuberculosis\u003c/em\u003e complex [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. \u003cem\u003eM. bovis\u003c/em\u003e zoonotically transmitted to humans; causing disease through ingestion; inhalation and less commonly by contact with mucous membranes and broken skin [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Bovine tuberculosis is characterized by the progressive development of granulomatous lesions (tubercles) in any body organ and can affect a broadly large number of species [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe intradermal tuberculin skin test has been in use for almost a century in the diagnosis of bovine tuberculosis and despite the technological advances of the last two decades, it is still the only reliable test for the diagnosis of tuberculosis in cattle [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The Purified Protein Derivative (PPD), the antigen used in the tuberculin test, contains many antigenic determinants of broad specificity, which leads to the appearance of non-specific reactors indicating false positive possibilities [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAlthough the tuberculin test is still considered the golden master test for diagnosis of bovine tuberculosis, it is tedious, laborious and, time-consuming. In addition, negative or doubtful tuberculin-tested cases cannot be retested before 6 months after tuberculin testing. Therefore, the recently developed serodiagnostic methods may be suitable and reasonable alternative tools for the diagnosis of bovine tuberculosis. Therefore, the suggestion of [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e] to use of ELISA with the tuberculin skin test to overcome the problems of the tuberculin test was very convincing. The first ELISA kit for the sero-diagnosis of bovine tuberculosis was introduced by [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], and it was for wildlife application using PPD-B as the target antigen. Previously developed ELISA kit was reported to have a sensitivity and specificity of 73% and 96%, respectively, for the detection of TB in wild boar [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs an alternative to tuberculin skin testing for antemortem diagnosis, quick and simple immune-chromatographic LFA assays for the serodiagnosis of bTB have been developed and presented. [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. As both qualitative captures and detectors of specific antibodies against \u003cem\u003eM. bovis\u003c/em\u003e in plasma, serum, and whole blood, these chromatographic immunoassays use special blends of chosen \u003cem\u003eM. bovis\u003c/em\u003e antigens. [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The common sero-reactive antigens in bovine TB have been identified as the MPB83, 14-kDa protein, ESAT-6, MPB70, CFP-10, MPT51, MPT32, MPB59, MPB64, MPT63, Acr1, PstS-1, 16-kDa alpha-crystalline/MPB83 fusion protein, ESAT-6/CFP10 fusion protein, \u003cem\u003eM. bovis\u003c/em\u003e purified protein derivatives, and \u003cem\u003eM. bovis\u003c/em\u003e culture filtrate [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The use of LFK in the sero-diagnosis of infectious diseases has been widely applied and its first application in the diagnosis of bovine tuberculosis was first recorded by [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Since then, these tests have become an extremely popular tool for the rapid diagnosis of bovine tuberculosis [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. However, the specificity and sensitivity of several commercially available LFA kits are questionable.\u003c/p\u003e \u003cp\u003eIn the present work, two LFKs were evaluated in the rapid detection of tuberculosis-infected cattle. The sensitivity and specificity of these kits were determined using the tuberculin skin test and bacterial isolation as a golden standard technique. The first kit was the IQRT Antigen Rapid Bovine TB antibody kit that employed recombinant \u003cem\u003eM. bovis\u003c/em\u003e MPB70 antigen (specific for \u003cem\u003eM. bovis\u003c/em\u003e) as capture and detector. The second kit was the Ubio quick VET Bovine TB Antibody Rapid Test, this kit uses a unique cocktail of recombinant antigens of \u003cem\u003eM. bovis\u003c/em\u003e to detect specific antibodies of three immunoglobulin classes, IgG, IgM, and IgA developed during infection in cattle.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Material And Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy design and animal population\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThrough a national study involving most of the Egyptian governorates, a total of 3750 crossbreed dairy cattle from different Egyptian Governorates, all over Egypt were involved in the current study. The study design can be clearly described through the flowchart illustrated in \u003cstrong\u003eFigure (1)\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eApplication of the single intradermal Tuberculin skin test done according to the\u0026nbsp;OIE manual (2009)\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;as follows;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTesting of each animal started with\u0026nbsp;measuring skin thickness\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003efollowed by an I/D injection 0.1 ml of bovine PPD.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eSeventy-two hours post-injection, the skin thickness was measured and the differences between the 1\u003csup\u003est\u003c/sup\u003e and 2\u003csup\u003end\u003c/sup\u003e reading in (mm) was recorded. According to the Egyptian General Organization of Veterinary Services \u003cstrong\u003e[15]\u003c/strong\u003e an increase in the skin thickness of 4mm or more was considered positive, less than 2mm is considered as negative and from 3- 4 mm was considered as doubtful. All positive reactants were slaughtered due to the \u0026ldquo;test and slaughter rule\u0026rdquo; in case of TB positive feedlot animal cases.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIsolation of acid-fast bacilli\u0026nbsp;from collected tissue samples of slaughtered tuberculin positive reactants:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tested tuberculin-positive cattle were slaughtered. Postmortem examination was conducted on them, and 69 tissue samples were collected. These samples were used for bacteriological isolation and identification of \u003cem\u003eM. bovis\u003c/em\u003e according to \u003cstrong\u003e[16]\u003c/strong\u003e.\u0026nbsp;Organs, lymph nodes, and/or other tissues were processed for the isolation of this organism. The collected sample processing was done as follows; all specimens were cut into small pieces, mixed with fine sterile sand, diluted with 2ml sterile distilled water, and ground in a sterile mortar. Two ml of 4% H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u0026nbsp;\u003c/sub\u003ewere added to the samples, which were then incubated for \u0026frac12; hour at 37\u003csup\u003eo\u003c/sup\u003eC. The mixture was furtherly diluted with 16 ml sterile distilled water and centrifuged at 3000 rpm/20 minutes. The obtained sediment was resuspended in 0.5 ml sterile distilled water, inoculated into Lowenstein-Jensen slants, and incubated at 37\u003csup\u003eo\u003c/sup\u003eC in an inclined position overnight and then vertically for at least 6-8 weeks with weekly examination starting from three days post inoculation and the obtained results were recorded \u003cstrong\u003e[17].\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSerological diagnosis of bovine tuberculosis using locally developed ELISA and 2 types of commercially available Lateral flow Kits:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll sero-diagnostic tests were performed on the serum samples that were collected from the examined dairy cattle before being tested with the single\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eintradermal\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003etuberculin skin test \u003cstrong\u003e[18]\u003c/strong\u003e. These samples were examined for diagnosis of bovine tuberculosis through the detection \u003cem\u003eMycobacterium bovis\u003c/em\u003e-specific antibodies. The serum samples were examined using a locally prepared ELISA and two commercially available lateral flow kits, namely; the Antigen Rapid Bovine TB Ab test Kit (IQRT test kits)\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eand\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ethe Ubio quick VET Bovine Tuberculosis Antibody Rapid\u0026nbsp;\u003c/strong\u003eTest\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eKit\u003c/strong\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe obtained results were comparably analyzed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Examination of the collected serum samples using the Anigen Rapid Bovine TB Ab test Kit (IQRT test kits) produced by Anigen Animal Genetics Inc., South Korea:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe test was performed per the manufacturer\u0026rsquo;s instructions as follows; three drops (about 60 - 90\u0026mu;l) of tested serum samples were dispensed into the sample application well. One drop of phosphate buffered saline was added into the sample well if the flow migration is not observed within 30 seconds in the result window, Results was read in 20 minutes with instructional invalid readings and interpretation after 20 minutes. Positive results appeared as two purple color bands (T band and C band) within the result window, positive results can be visible in as short as 1 minute. If the intensity of the purple band color is faint, it is interpreted as positive as it appears within 20 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Examination of the collected serum samples using the \u003cstrong\u003eUbio quick VET Bovine Tuberculosis Antibody Rapid\u0026nbsp;\u003c/strong\u003eTest\u003cstrong\u003e\u0026nbsp;Kit produced by Ubio, India:\u003c/strong\u003e\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe test was performed per the manufacturer\u0026rsquo;s instructions as follows; three drops (about 60 - 90\u0026mu;l) of the tested serum were added to the sample hole on the test card using a dropper. The result was recorded in 5-10 minutes. The control line, marked C, should \u0026nbsp; always develop color if the test procedure is performed properly and the test reagents are working. A red test line marked T will be visible if there are enough Bovine TB specific antibodies in the serum sample.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Examination of the collected serum samples using locally prepared\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eELISA:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe antigen used for coating the microtiter plate wells was bovine PPD (B-PPD) that was obtained from the Bacteriological Diagnostic Products Department, Veterinary Serum and Vaccine Research Institute (VSVRI), Abbasia, Cairo, Egypt. It was prepared according to the protocol of the Central Veterinary Laboratories, Weighbridge, United Kingdom. Briefly, the test was developed as follows; Polystyrene 96-well microtiter plates were coated with the B-PPD antigen using 15\u0026micro;g/ml in 0.1M carbonate-bicarbonate buffer (pH 9.6). The antigen solution was dispensed in 50ul/well. In each microtiter plate 4 wells were left uncoated with B-PPD antigen and served as blank control wells. Each of these blank wells received 50\u0026micro;l of carbodiimide in 0.1 M carbonate bicarbonate coating buffer. The plates were incubated overnight at 4\u0026deg;C then decanted, washed 3 times with PBS and air dried for 10 minutes. Then all wells except the blank ones were filled 100\u0026micro;l/well of 0.01 M ammonium chloride as blocking buffer and further incubated for 30 minutes at 22\u0026ordm;C. Finally, the plates were washed 3 times with PBS washing buffer \u003cstrong\u003e[19]\u003c/strong\u003e. In each microtiter plate, all wells except the wells of the first column were filled with 50ul/well PBS buffer. For each serum sample one row is used. The tested serum samples were diluted 1:40 in PBS and 100\u0026micro;l of each tested serum sample was added to the first well of the corresponding row. This was followed by two-fold serial dilutions of each sample. Each plate received negative serum samples run in the same dilutions to calculate the cut-off value. The plates were incubated at room temperature for 30 minutes on a horizontal shaker. The microtiter plates were decanted, washed 4 times with ELISA washing buffer and allowed to stand inverted for 30minutes. To each well, 50\u0026micro;l of anti-bovine IgG alkaline phosphatase diluted 1:5000 in diluting buffer were added and the plates were then incubated for 30 minutes at room temperature on a horizontal shaker. The plates were again washed 4 times with ELISA washing buffer and allowed to stand inverted for 30 minutes. The substrate solution composed of P- Nitrate phenyl phosphate (one tablet 5mg /5ml substrate buffer) was prepared and 50\u0026micro;l/well was dispensed in all wells of the plates and were further incubated for 20min at room temperature. The color development was stopped by the addition of 100\u0026micro;l/well of 0.4 M NaOH-stopping solution. The results were recorded by optical density measurement at 405 nm using spectra III ELISA reader. A titer of 1:80 or higher was considered positive for bovine tuberculosis \u003cstrong\u003e[20].\u0026nbsp;\u003c/strong\u003eThe results obtained by the applied serological tests were compared with the tuberculin testing and the results of the culture isolation, which is considered the gold standard test. The diagnostic value of the tested serological methods was determined in the terms of accuracy index, sensitivity, specificity, positive predictive value, and negative predictive value. \u0026nbsp;\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003eBovine tuberculosis (bTB) is a zoonotic disease with public health significance. The maximum detection of bovine TB in cattle populations in Egypt is vital to understand its epidemiology and zoonotic potentials and to achieve significant reduction and control of the disease in livestock. \u0026nbsp;The tuberculin skin tests (TST) are currently the best available techniques for international field diagnosis of bTB in live animals \u003cstrong\u003e[2, 21]\u003c/strong\u003e and it is based on delayed hypersensitivity reactions. \u0026nbsp;The results of tuberculin skin testing are presented in In \u003cstrong\u003eTable (1)\u003c/strong\u003e where out of 3750 tuberculin-tested cross-bred dairy cattle, 69 were found to be positive reactors with a prevalence rate of 1.8%.\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"582\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFarm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal no. of examined animals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"36.25429553264605%\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive tuberculin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"46.44549763033175%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"53.55450236966825%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e170\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e220\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e290\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e140\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e320\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e500\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e12\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e410\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e250\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e180\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e290\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e370\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e12\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e155\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e225\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e14\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e110\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003e15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e120\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.83\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"28.694158075601376%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.05154639175258%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3750\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.83848797250859%\"\u003e\n \u003cp\u003e\u003cstrong\u003e69\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.415807560137456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.8\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\u003cstrong\u003eTable (1):\u0026nbsp;\u003c/strong\u003eTuberculin skin test result in dairy farm cattle from some Egyptian Governorates.\u003c/p\u003e\n\u003cp\u003eThis is comparatively lower than that recorded by other investigators in Egypt \u003cstrong\u003e[22], (6.9%), [23],\u003c/strong\u003e (\u003cstrong\u003e4.6%), and [24], (2.2%)\u003c/strong\u003e and in some other African countries as\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ein Ethiopia\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(11.6%) \u003cstrong\u003e[25],\u0026nbsp;\u003c/strong\u003eand in Chad (8%) \u003cstrong\u003e[26]\u003c/strong\u003e. This noticable reduced incidence might be attributed to the fact that the farms included in this research perform the tuberculin test regularly and applied test and slaughter strategy \u003cstrong\u003e[27]\u003c/strong\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eOn the other hand, the prevalence rate recorded in the present study is comparatively higher than that reported by other investigators in other countries, 0.9% to 1.3% in Tanzania\u003cstrong\u003e\u0026nbsp;[28, 29],\u0026nbsp;\u003c/strong\u003erespectively, 0.54 % in Venezuela \u003cstrong\u003e[30]\u003c/strong\u003e, 0.4 %, in New Zealand\u003cstrong\u003e\u0026nbsp;[31]\u0026nbsp;\u003c/strong\u003e0.05 to 0.15 % in Australi\u003cstrong\u003ea [32]\u003c/strong\u003e and\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e0.02 % in Japan\u003cstrong\u003e\u0026nbsp;[33]\u003c/strong\u003e. The postmortem findings of the slaughtered TB reactors revealed that 73.9% of these animals showed visible lesions. At the same time the percentage of tuberculin reactors with non-visible lesions were 26.1%, \u003cstrong\u003eTable (2)\u003c/strong\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"595\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal No. of animals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"10\" width=\"72.94117647058823%\"\u003e\n \u003cp\u003e\u003cstrong\u003eVisible lesions\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNon- visible l\u0026eacute;sions\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" width=\"21.839080459770116%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeneralized\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"19.54022988505747%\"\u003e\n \u003cp\u003e\u003cstrong\u003eHead\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"19.54022988505747%\"\u003e\n \u003cp\u003e\u003cstrong\u003ePulmonary\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"19.54022988505747%\"\u003e\n \u003cp\u003e\u003cstrong\u003eDigestive\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"19.54022988505747%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMixed\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" width=\"12.794612794612794%\"\u003e\n \u003cp\u003e\u003cstrong\u003e69\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.228956228956229%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.080808080808081%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.228956228956229%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.397306397306397%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.397306397306397%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.397306397306397%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.912457912457913%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e11.6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.142857142857143%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.266409266409266%\"\u003e\n \u003cp\u003e\u003cstrong\u003e7.2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.142857142857143%\"\u003e\n \u003cp\u003e\u003cstrong\u003e22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e31.9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.335907335907336%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8.7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.335907335907336%\"\u003e\n \u003cp\u003e\u003cstrong\u003e14.5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.335907335907336%\"\u003e\n \u003cp\u003e\u003cstrong\u003e18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.073359073359073%\"\u003e\n \u003cp\u003e\u003cstrong\u003e26.1\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\u003cstrong\u003eTable (2):\u003c/strong\u003e Result of Postmortem examination of tuberculin positive slaughtered cattle.\u003c/p\u003e\n\u003cp\u003eThese results agreed with those reported by \u003cstrong\u003e[34, 35]\u003c/strong\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eOn the other hand, a much higher percentage was recorded by \u003cstrong\u003e[36]\u003c/strong\u003e who reported (86.9%) in Queensland, \u003cstrong\u003e[37]. \u0026nbsp;\u0026nbsp;\u003c/strong\u003eIn Egypt Lower percentage was also reported by \u003cstrong\u003e[38],\u0026nbsp;\u003c/strong\u003e(46%),\u003cstrong\u003e\u0026nbsp;[39],\u003c/strong\u003e (52.7%), and\u003cstrong\u003e\u0026nbsp;[40],\u0026nbsp;\u003c/strong\u003e(40.8%). The percentage of tuberculin positive animals with non-visible lesions was attributed to sensitization by atypical mycobacteria or even closely related microorganisms especially the members of the genus Nocardia or a combination of Liver Fluke infestation with saprophytic mycobacteria \u003cstrong\u003e[41, 42].\u0026nbsp;\u003c/strong\u003eMoreover, \u003cstrong\u003e[43, 44]\u0026nbsp;\u003c/strong\u003eattributed the cause of non-specific reaction to the assumption that, those animals might be slaughtered at early stage of the disease where the tuberculous lesions were still invisible, or lesions may be found in parts of the body not routinely examined in the carcass such as bone or brain. The relation between the postmortem findings in different ages of tuberculin reactor cattle is presented in \u003cstrong\u003eTable (3).\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e \u003cimg width=\"624\" 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\" alt=\"image\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable (3):\u003c/strong\u003e Results of correlation between age of animal and PM finding of the slaughtered cases\u003c/p\u003e\n\u003cp\u003eThe percentage of reaction-positive animals increased with age, reaching a maximum in animals over 60 months old. Similar studies in Great Britain recorded an increase of incidence of bTB with increased age \u003cstrong\u003e[45]\u003c/strong\u003e. It was suggested earlier by \u003cstrong\u003e[46]\u0026nbsp;\u003c/strong\u003ethat, the high positivity of the old age might be due to the reduced number \u0026gamma; and \u0026delta; T cells responsible of the anti-mycobacterial immunity. [These cells are predominantly found in the circulation of young calves. It has also been suggested that increased incidence of TB in older animals can be due to a waning of protective capability in aged animals \u003cstrong\u003e[47, 48]\u003c/strong\u003e. The correlation between site of lesion and the isolated mycobacteria species is shown in \u003cstrong\u003eTable (4).\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"632\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" width=\"33.06962025316456%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eNo. of processed \u0026nbsp;tissue samples\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"66.93037974683544%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Bacteriological isolation (52 isolates)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"33.096926713947994%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eM. bovis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"66.903073286052%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Unidentified mycobacteria\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"17.92452830188679%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"33.490566037735846%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSlow growers\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"33.490566037735846%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRapid growers\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.760563380281692%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.239436619718308%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.04225352112676%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.95774647887324%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.0173775671406%\"\u003e\n \u003cp\u003e\u003cstrong\u003e69\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.110584518167457%\"\u003e\n \u003cp\u003e\u003cstrong\u003e48\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.006319115323855%\"\u003e\n \u003cp\u003e\u003cstrong\u003e69.6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.006319115323855%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.42654028436019%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.375987361769353%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.056872037914692%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.4\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\u003cstrong\u003eTable (4):\u003c/strong\u003e Result of bacteriological isolation and identification of mycobacteria isolated from tuberculin positive cattle. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe total acid-fast bacilli isolated from the 69 slaughtered cattle were 52 isolates (75.4%). According to the morphological characters, growth rate, pigmentation, growth at different temperatures and biochemical tests these isolates were identified into 48\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003e(69.6%) \u003cem\u003eM. bovis\u003c/em\u003e, 1 unidentified rapid grower strain (1.44%) and 3 (4.3%) unidentified pigmented slow grower strains. The recovery rate of \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003efigured up to 69.6% but other authors reported a lower \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003erecovery rates, 14.8% by \u003cstrong\u003e[49],\u003c/strong\u003e\u0026nbsp; 20.2% by \u003cstrong\u003e[50],\u003c/strong\u003e\u0026nbsp; 41% \u0026nbsp;by \u003cstrong\u003e[51\u003c/strong\u003e], 35.4% by \u003cstrong\u003e[52],\u003c/strong\u003e 29.1% by \u003cstrong\u003e[53]\u003c/strong\u003e\u003cem\u003e,\u003c/em\u003e 5.6% by\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e[54]\u003c/strong\u003e, and 42.9% by \u003cstrong\u003e[55]\u003c/strong\u003e. On the contrary, \u003cstrong\u003e[56]\u003c/strong\u003e, in \u0026nbsp;Korea \u0026nbsp; reported \u0026nbsp;a \u0026nbsp;much \u0026nbsp; higher \u0026nbsp;isolation \u0026nbsp;rate (92.1%), in the republic of Korea. \u0026nbsp;These results depend mainly on the actual disease status present in the tested herd and to some extent on the experience of the investigators as well as the technique used for decontamination of tissue specimens. Data presented in\u003cstrong\u003e\u0026nbsp;Figure (2)\u003c/strong\u003e, shows the relation between the post-slaughter findings of positive tuberculin reactors and the type of mycobacteria recovered from the 69 carcasses,\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003eM. bovis isolates\u003cstrong\u003e,\u003c/strong\u003e\u003c/em\u003e rapid growers isolates and slow growers pigmented isolates were recovered at rates of 69.6 %,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e4.3%, and 1.44 %, respectively. \u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003eM. bovis\u003c/em\u003e was isolated at a rate of 100% from generalized TB, 91% from pulmonary TB, 81.8% from Extra Pulmonary TB\u003cstrong\u003e,\u0026nbsp;\u003c/strong\u003e90% from mixed TB, and 11.1% from non-visible lesion (NVL) reactors. These results coincide with those reported by \u003cstrong\u003e[57]\u003c/strong\u003e who isolated \u003cem\u003eM. bovis\u003c/em\u003e from NVL reactors. On the other hand, the recovery rate of typical mycobacteria rapid growers and slow growers pigmented was 16.7% of the extra-pulmonary for both of them and 33.3% of the mixed for the rapid growers. None of the cases of generalized tuberculosis yielded atypical mycobacteria, since \u003cem\u003eM.\u003c/em\u003e\u003cem\u003e\u0026nbsp;bovis\u003c/em\u003e has a tendency to induce progressive lesions and generalization \u003cstrong\u003e[44, 58]\u003c/strong\u003e. All cases with generalized TB yielded \u003cem\u003eM. bovis, a\u003c/em\u003e finding indicating the high susceptibility of cattle to \u003cem\u003eM. bovis\u003c/em\u003e infection. Comparison between bacteriological examination and serological testing by lateral flow kits of tuberculin-positive cattle are shown in \u003cstrong\u003eFigure (3)\u003c/strong\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe results of the application of lateral flow kits on sera from tuberculin-positive cattle\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eThe One- step Anigen Rapid Bovine Antibody Test\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(IQRT) employed recombinant \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003eMPB70 antigen specific for \u003cem\u003eM. bovis\u003c/em\u003e as capture and detector antigen. It is important to also mention that serological testing was conducted prior to TST because the latter can boost antibody responses in infected cattle and emphasizes the importance of timing of collection of blood samples on the interpretation the test \u003cstrong\u003e[59].\u0026nbsp;\u003c/strong\u003eThis Anigen TB antibody test kit has detected 43.8% of tuberculin positive cattle against 69.6% that were confirmed by bacterial isolation of \u003cem\u003eM. bovis\u003c/em\u003e. While the other lateral flow assay \u003cstrong\u003eUbio quick VET\u0026nbsp;\u003c/strong\u003etest is a single-directional lateral-flow serological test, which was produced to provide a quick determination of the presence of \u003cem\u003eM. bovis\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eantibody \u003cstrong\u003e[60, 61]\u003c/strong\u003e has detected zero % of tuberculin positive cattle. Comparison between the result of ELISA test, rapid lateral flow test and bacteriological isolation of tuberculin positive cattle showed that 8 tuberculin positive reactors with generalized tuberculous lesions were bacteriologically positive\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e(M. bovis\u0026nbsp;\u003c/em\u003eisolation) and ELSIA positive using bovine PPD as coating antigen (100%).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAs shown in \u003cstrong\u003eFigure (4)\u003c/strong\u003e, out of 22 tuberculin positive reactors 10 (45.5%) showed pulmonary lesion, out of 11 tuberculin positive reactors 3 (27.3%) showed extra-pulmonary lesions (head and digestive), and Out of 10 tuberculin reactors showing mixed TB lesions (pulmonary and extra pulmonary), 5 (50%) isolates of \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003ewere positive for ELISA by using bovine PPD as coating antigens. Out of 18 tuberculin reactors showing NVL, 2 (11.1%) isolates of \u003cem\u003eM. bovis\u003c/em\u003e could be isolated from them and zero (0%) serum samples were positive by ELISA by using bovine PPD as coating antigen.\u003c/p\u003e\n\u003cp\u003eThe results obtained in the current study agreed with those reported by \u003cstrong\u003e[62, 63]\u0026nbsp;\u003c/strong\u003ewho stated that, antibody response to \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003einfection is certainly not uniform. This phenomenon was previously suggested in serological analysis by \u003cstrong\u003e[64]\u003c/strong\u003e who studied the humoral immune response to \u003cem\u003eM. bovis\u0026nbsp;\u003c/em\u003einfection in cattle and concluded that, it was characterized by highly heterogeneous antigen recognition. It is concluded that single intradermal tuberculin test, culture and isolation of the mycobacterium are recommended for determination of a true prevalence of bTB in the herd to be established.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe recent lateral flow rapid kits could be used for initial tuberculosis screening in combination with tuberculin skin test (TST) for improving sensitivity of bovine tuberculosis screening, thereby leading to more successful control programs in developing countries. The rapid test is proposed as a potentially useful ancillary assay for bTB. In addition, it may be most suitable for surveillance, especially if an immediate result is needed. The differences in the sensitivity and specificity of the different LFA kits sources available in the market necessitate that, the importance of choosing type of diagnostic kits used for rapid detection of bovine tuberculosis. Also, these kits alone may be not enough for accurate diagnosis of bovine tuberculosis.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe current conducted study is reported in accordance with (Animal Research: Reporting of In-Vivo Experiments-ARRIVE) guidelines. The guidelines of the (Institutional Animal Care and Use Committee-IACUC of faculty of veterinary medicine, Cairo University) were completely followed during any procedures involving animal use through the current conducted study. This study is approved by the ethical committee of the institutional animal care and use committee (IACUC) of Veterinary-Cairo University. \u003cem\u003e\u003cu\u003e\u0026nbsp;\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eNo anesthesia or euthanasia protocols were used with the animal involved during this study as all animal-dependent methodological procedures were considered as no to low pain-causing procedures that ethically can be done on a conscious alive animal.\u003c/p\u003e\n\u003cp\u003eAll sample collection procedures were done based on permission from the cattle farm owners and managers.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026apos;Not applicable\u0026apos;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed related to the animal cases tested during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026apos;Not applicable\u0026apos;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed equally during the whole stages of the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank all general practitioner veterinarians and dairy animal specialists all over Egypt due to their help during the sample collection stages.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eTaylor GM, Worth DR, Palmer S, Jahans K, Hewinson RG. 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Vet Microbiol. 1992;31:243\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Bovine tuberculosis, tuberculin skin test, M. bovis isolation, lateral flow kits for antibody detection, ELISA, Rapid diagnostics","lastPublishedDoi":"10.21203/rs.3.rs-2121571/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2121571/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBovine tuberculosis is a worldwide disease that causes great economic losses to the dairy industry and constitutes a serious human public health hazard. In the present study, two types of commercially available immunochromatographic lateral flow kits, namely, IQRT Anigen Rapid Bovine TB Ab kit and Ubio quick VET Bovine Tuberculosis Antibody kit were evaluated for their sero-diagnostic sensitivity and specificity of bovine tuberculosis through the detection of \u003cem\u003eMycobacterium bovis\u003c/em\u003e-specific antibodies in tested serum samples. The results obtained by these kits were compared with the ELISA test, bacteriological examination, and tuberculin skin testing. In this study, out of 3750 dairy cattle tested by single intradermal tuberculin skin test, 69 (1.8%) animals reacted positively. From the slaughtered tuberculin-positive cattle 51 animals (73.9%) showed visible lesions on postmortem examination while the remaining 18 animals (26.1%) showed no visible lesions. Using bacteriological examination of tissue specimens collected, 48 out of the 69 positive tuberculin reactor animals (69.6%) were positive and \u003cem\u003eM. bovis\u003c/em\u003e was successfully isolated. Using ELISA for serological examination of serum samples collected prior to skin testing from the 69 positive tuberculin reactors, only 26 cases (37.7%) were positive. The serodiagnosis of bovine tuberculosis using the IQRT Anigen lateral flow kit, which employed recombinant \u003cem\u003eM. bovis\u003c/em\u003e MPB70 antigen as capture and detector, could identify only 21 positive cases, which represent 43.8% of 48 bacteriologically identified cases and 30.44% out of the 69 tuberculin positive cases. On the other hand, the Ubio quick VET kit has detected zero% of bovine tuberculosis-positive cattle. It is concluded that the use bovine tuberculosis Antibody Rapid Test Kit alone may be unreliable for the detection of tuberculosis-infected cattle.\u003c/p\u003e","manuscriptTitle":"A critical appraisal on antibody detector-serodiagnostics for Bovine Tuberculosis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-10-14 17:39:51","doi":"10.21203/rs.3.rs-2121571/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":"8884f604-c763-4eb6-9025-616e796ed8a8","owner":[],"postedDate":"October 14th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-11-16T10:29:13+00:00","versionOfRecord":[],"versionCreatedAt":"2022-10-14 17:39:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2121571","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2121571","identity":"rs-2121571","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}