Optimization of the Fecal Pool Method for the Detection of <i>Mycobacterium avium </i>Subspecies <i>paratuberculocis </i>with Quantitative Real-Time PCR

DOI JASI Web Site Web Site 3 References

Bibliographic Information

Other Title
  • ヨーネ病リアルタイムPCR検査における糞便のプール処理法の検討
  • ヨーネビョウ リアルタイム PCR ケンサ ニ オケル フンベン ノ プール ショリホウ ノ ケントウ

Search this article

Abstract

The objective of this study was to obtain the optimal fecal pool method for the screening of Mycobacterium avium subspecies paratuberculosis (Map) infection with quantitative real-time PCR (qPCR). Initially, the effect of pooling on the recovery rate of Map DNA was evaluated using a fecal sample from experimentally Map-infected cattle. Map DNA concentrations extracted and purified from pooled fecal samples were increased in proportion to the volume of feces, and almost all Map DNA was recovered without any PCR inhibitory effect by increasing fecal volume. In the qPCR test at a pooling rate of 1 infected plus 2 to 5 uninfected cattle, all pooled samples were qPCR positive, and the calculated Map DNA concentrations of 0.0022 to 0.0024 pg/PCR from pooled samples were almost the same as that of one infected cattle up to 5pool (1 infected plus 4 uninfected cattle). These results suggest that the sensitivity of qPCR is not affected by fecal sample pooling up to 5pool. This pooling and centrifugation method seems likely to be useful for herd screening and improving the sensitivity of the Map qPCR test.

Journal

References(3)*help

See more

Keywords

Details 詳細情報について

Report a problem

Back to top