Evaluation of real time PCR for the detection of Mycobacterium avium subsp. paratuberculosis in faecal samples of cattle

Abstract

The efficiency and suitability of a MAP F57 based SYBR Green qPCR assay for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) using a set of known MAP positive (12) and MAP negative (23) DNA samples that were previously identified by conventional IS 900 PCR were assessed. These DNA samples were isolated in our previous study from faecal samples collected from cattle in the livestock farms under government sector with a previous history of Johne’s disease. The MAP F57 qPCR was able to identify all the positive samples accurately and rapidly with Cq values ranging from 20-29.The efficiency of qPCR using recombinant plasmid for standard curve was 0.991 and limit of detection was 10 MAP organisms per microlitre of DNA sample.

Keywords

• johne’s disease
• map
• qpcr
• f57