Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
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Multiplex Quantitative Real-time Polymerase Chain Reaction Assay for Rapid Detection of Mycobacterium avium subsp. paratuberculosis in Fecal Samples

분변 시료에서 Mycobacterium avium subsp. paratuberculosis 의 빠른 검출을 위한 다중 실시간 중합효소연쇄반응기법의 개발

  • Accepted : 2015.05.18
  • Published : 2015.06.30
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Abstract

Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis or Johne's disease, an intestinal granulomatous infection in domestic and wild animals. The study aimed to develop and evaluate a panel of multiplex quantitative real-time polymerase chain reaction (mqPCR) assay for simultaneous detection of three MAP-specific genes (IS900, F57 and ISMAP02 genes). The analytic sensitivity (i.e., limit of detection, expressed as cells per 1 ml) was 150 for IS900, 1500 for F57, and 50 for ISMAP02. The specificity of the method was determined by testing 152 bovine fecal samples. Based on the test, it showed that the assay simultaneously detected the target genes in short period of time and at lower cost compared to laboratory routine tests. The test agreement between the assay and routine test was 94%. The discrepancy in the results was due to samples that were tested positive by the panel but negative by the routine tests, suggesting that the assay has higher sensitivity than the routine tests. In conclusion, the mqPCR assay could be a rapid and accurate testing tool for investigating paratuberculosis or Johne's disease cases in domestic and wild animals.

Mycobacterium avium subsp. paratuberculosis (MAP)는 가축 및 야생동물에서 장 내 육아종성 감염증을 유발한다. 이 연구의 목적은 MAP 특이유전자 3개(IS900, F57 및 ISMAP02)의 빠른 검사를 위한 다중 실시간 중합효소연쇄반응기법을 개발하고 평가하는데 있다. 평가 결과 분석 민감도는 IS900이 150 cells/ml, F57이 1500 cells/ml, ISMAP02가 50 cells/ml로 확인되었다. 152개 소 분변시료를 대상으로 실시한 검사 결과 개발한 기법이 기존 검사방법보다 빠르고 적은 비용으로 동시검사가 가능한 것으로 확인되었다. 검사결과의 일치도는 94%로 나타났다. 불일치 결과는 개발한 기법이 양성으로 확인하였으나, 기존 검사에서는 음성으로 나타난 것 때문으로 확인되어, 개발한 기법이 더 높은 민감도를 갖는 것으로 나타났다. 개발한 다중 실시간 중합효소연쇄반응기법은 가축 및 야생동물에서 파라결핵 또는 요네병의 조사를 위한 빠르고 정확한 검사기법이 될 것이다.

Keywords

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