등온증폭법을 이용한 고감도 JC polyomaviruses 진단법 개발
Diagnostic Method for the Detection of JC Polyomavirus Using Loop-mediated Isothermal Amplification
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초록
JC polyomavirus (JCPyV)는 viral group I, Polyomaviridae로분류되는 사람 병원성 바이러스 이다. JCPyV는최근 하수 등 수질오염 지표로 제안됨에 따라 수질환경에서 모니터링 필요성이 제기되고 있다. 임상 및 환경 시료 중 JCPyV 검출을 위해서 PCR 시스템이 사용되어 왔다. 그러나 신속하고 높은 검출 민감도를 가진 방법의 필요성에 따라 이번 연구에서는등온증폭 프라이머 조합을 개발하였다. 이번에 개발한 방법은 기존 PCR 시스템에 비해 더욱 신속 및 약 10배 더 높은 검출 민감도를 보여주었다. 또한 본 연구에서 개발한 검출 방법의 재 검정을 위해서 HaeIII 제한효소를 이용한 방법을 함께 고안하였다. 따라서 이번 연구는 임상, 환경 등의 시료에서 JCPyV를 모니터링 하기 위한 방법으로 활용이 기대된다.
Abstract
JC polyomavirus (JCPyV) is a human pathogenic virus belonging to the family Polyomaviridae, a viral group containing dsDNA nucleic acid. A recent recommendation is to apply the presence of JCPyV as a fecal indicator for water contamination in environments like sewage, and techniques to monitor JCPyV in water are being proposed. To date, the conventional PCR system has been applied as a diagnostic method for detecting JCPyV. There is a need for a more rapid and sensitive JCPyV diagnostic detection method in clinical and environmental samples. In this study, we developed a loop-mediated isothermal amplification (LAMP) primer set for the detection of JCPyV. Our results indicate that the LAMP method using a specific primer set shows about 10-fold higher detection sensitivity than the conventional PCR system. The effectiveness of the LAMP method developed in this study has been validated by PCR product digestion using the HaeIII restriction enzyme. We, therefore, propose that the LAMP method using a specific primer set can be applied as a rapid and sensitive detection method for monitoring JCPyV in clinical and environmental samples.
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