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.

Abstract

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.

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