Polymerase spiral reaction assay for rapid visual detection of Mycobacterium avium subsp. paratuberculosis in fecal samples.

ABSTRACT: A rapid, sensitive, and specific visual detection assay for Mycobacterium avium subsp. paratuberculosis (MAP) was developed and optimized using a polymerase spiral reaction (PSR) method. A pair of primers was designed targeting MAP specific sequence of IS900 putative transposes (p43) gene, and PSR results were assessed using agarose gel electrophoresis and colour change with SYBR Green-I dye. The assay was optimized using water bath and the optimum reaction time and temperature for MAP-PSR were 60 min and 64 °C, respectively. The sign of target amplification can be visualized by naked eyes as SYBR Green-I change its colour due to intercalation with amplified products. The developed assay demonstrated that the primers specifically detected MAP and showed no cross-reaction with other common

SUBMITTER: Singh VK 

PROVIDER: S-EPMC12297387 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature