35-Title: Molecular identification of Mycobacterium avium subspecies paratuberculosis by using conventional PCR
Authors: Gaurav Agrawal, DS Meena, Sandeep Kumar Sharma, CS Sarswat, Nazeer Mohammed and Dilip Singh Meena
Source: Ruminant Science (2022)-11(2):431-434.
How to cite this manuscript: Agrawal Gaurav, Meena DS, Sharma Sandeep Kumar, Sarswat CS, Mohammed Nazeer and Meena Dilip Singh (2022). Molecular identification of Mycobacterium avium subspecies paratuberculosis by using conventional PCR. Ruminant Science 11(2):431-434.
The present study was conducted for molecular confirmation of insertion sequence, IS900 of Mycobacterium avium subspecies paratuberculosis (MAP) by using conventional PCR. A total of 200 blood samples from milking cattle were collected from the nearby area of PGIVER Institute, Jaipur, Rajasthan. Total Genomic DNA was extracted from whole blood (EDTA). PCR amplification was performed on DNA extracted from collected blood samples for the presence of insertion sequence IS900 of MAP. The samples were considered positive at 707 bp PCR amplified band was present. PCR products were visualized by 1.5% agarose gel. Out of 200 blood samples, only 21 (10.5%) samples were found to be positive in PCR. Therefore, the prevalence recorded for MAP based on molecular tools is 10.5%. In conclusion, PCR targeting the IS900 element has shown high specificity for MAP, a fact that minimizes the risk of false negative and positive results.
Bhati Anita, Shringi Archana, Vyas Indu, Mandovera Vijay and Rani Sunita (2013). A study of pathological conditions in buffalo intestine. Ruminant Science 2(1):43-44.
Bhide M, Chakurkar E, Tkacikova L, Barbuddhe S, Novak M and Mikula I (2006). IS900-PCR-based detection and characterization of Mycobacterium avium subsp. paratuberculosis from buffy coat of cattle and sheep. Veterinary Microbiology 112(1):33-41.
Buergelt CD and Williams JE (2004). Nested PCR on blood and milk for the detection of Mycobacterium avium subsp paratuberculosis DNA in clinical and subclinical bovine paratuberculosis. Australian Veterinary Journal 82(8):497-503.
Harris NB and Barletta RG (2001). Mycobacterium avium subsp. paratuberculosis in veterinary medicine. Clinical Microbiology Reviews 14(3):489-512.
Hendrick SH, Kelton DF, Leslie KE, Lissemore KD, Archambault M and Duffield TF (2005). Effect of paratuberculosis on culling, milk production, and milk quality in dairy herds. Journal of the American Veterinary Medical Association 227(8):1302-1308.
Ikonomopoulos J, Gazouli M, Pavlik I, Bartos M, Zacharatos P, Xylouri E, Papalambros E and GorgoulisV(2004). Comparative evaluation of PCR assays for the robust molecular detection of Mycobacterium avium subsp. paratuberculosis. Journal of Microbiological Methods 56(3):315-321.
Lu Z, Mitchell RM, Smith RL, Van Kessel JS, Chapagain PP, Schukken YH and Grohn YT (2008). The importance of culling in Johne’s disease control. Journal of Theoretical Biology 25(1):135-146.
Mohan A, Das P, Kushwaha N, Karthik K and Niranjan AK (2013). Investigation on the status of Johne’s disease based on agar gel immuno diffusion, Ziehl-Neelsen staining and nested PCR approach in two cattle farm. Veterinary World 6(10):778-784.
Park HT, Shin MK, Park HE, Cho YI and Yoo HS (2016). PCR-based detection of Mycobacterium avium subsp. paratuberculosis infection in cattle in South Korea using fecal samples. Journal of Veterinary Medical Sciences 78(9):1537-1540.
Rani M, Narang D, Kumar D, Chandra M, Singh ST and Filia G (2018). IS Map02 element targeted nested polymerase chain in the detection of Mycobacterium avium subsp. paratuberculosis in fecal samples of cattle and buffaloes. Veterinary World 11(3):397-401.
Sambrook JF and Russell DW (2001). Molecular cloning: a laboratory manual. 3rd Edn, Cold Spring Harbor Laboratory, New York, USA, p 112.
Sahzad, Gupta Saurabh, Chaubey Kundan Kumar, Jayaraman Sujatha, Singh Manju, Stephan Bjorn John, Dayal Deen, Jain Mukta, Pachoori Anjali, Sohal Jagdip Singh, Bhatia Ashok Kumar and Singh Shoor Vir (2015). Comparative evaluation of two screening tests (serum elisa and fecal microscopy) for the estimation of Johne’s disease infection in goat herds endemically infected with Johne’s disease. Ruminant Science 4(2):171-174.
Singh SV, Singh PK, Singh AV, Sohal JS, Kumar N, Chaubey KK, Gupta S, Rawat KD, Kumar A, Bhatia AK and Srivastav AK (2014). ‘Bio Load’and Bio Type profiles of Mycobacterium avium subspecies paratuberculosis infection in the domestic livestock population endemic for Johne’s disease: A survey of 28 years (1985-2013) in India. Transboundary and Emerging Diseases 61:43-55.
Speksnijder DC, de Wit MK and Krattley Roodenburg B (2022). Detection of a high prevalence of paratuberculosis in a previously test negative conventional dairy herd in the Netherlands. Veterinary Record Case Reports 10:e290.
Tiwari Rahul, Singh Rashmi, Jeph Nirmal K, Meena DS, Mohammad Nazeer and Meena Sarjna (2021). Prevalence of paratuberculosis in cattle of Jaipur city, Rajasthan. Ruminant Science 10(2):383-388.
Uy MRD, Cruz JL, Miguel MA, Salinas MBS, Lazaro JV and Mingala CN (2018). Serological and molecular evaluation of Mycobacterium avium subspecies paratuberculosis (Johne’s disease) infecting riverine-type water buffaloes (Bubalus bubalis) in the Philippines. Comparative Immunology, Microbiology and Infectious Disease 61:24-29.
Whittington RJ and Sergeant ESG (2001). Progress towards understanding the spread, detection and control of Mycobacterium avium subsp para tuberculosis in animal populations. Australian Veterinary Journal 79(4):267-278.
Wolf R, Orsel K, De Buck J and Barkema HW (2015). Calves shedding Mycobacterium avium subspecies paratuberculosis are common on infected dairy farms. Veterinary Research 46(1):1-8.