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경남지역 종합병원에서 분리된 그람음성막대균으로부터 blaKPC 및blaNDM 유전자 검출

Detection of blaKPC and blaNDM Genes from Gram-Negative Rod Bacteria Isolated from a General Hospital in Gyeongnam

박지애|양병선
대한임상검사과학회 (Korean Society for Clinical Laboratory Science)|2021.03|대한임상검사과학회지 (Korean Journal of Clinical Laboratory Science)|제53권 제1호|pp. 49-59|DOI 10.15324/kjcls.2021.53.1.49

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키워드

MHTReal-time PCRCITblaKPCblaNDM
주제분류기타자연과학

초록

본 연구는 국내에서 가장 빈번히 검출되는 CPE의 유전자형중 blaKPC 및 blaNDM 유전자의 진단으로 기존의 표현형 검사 및일반 PCR 검사보다 시간 단축 및 사후 분석의 단점이 보완된real-time PCR의 융해 곡선을 이용한 분석법에 대해 알아보았다. 표현형적 검사결과 MHT는 35균주 중 25균주에서 양성을확인하고, CIT는 meropenem+PBA 및 meropenem+EDTA 에서 각각 14균주의 양성을 확인하였다. PCR 검사결과 KPC 25균주에서 증폭 산물을 확인하였고, K. pneumoniae 10균주, E. coli 5균주, A. baumannii 5균주, P. aeruginosa 4균주, P. putida 1균주로 나타났다. NDM은 8균주에서 증폭 산물을 확인하였고, K. pneumoniae 2균주, E. coli 3균주, P. aeruginosa 1균주, E. cloacae 1균주, P. rettgeri 1균주로 나타났다. 실시간 중합효소연쇄반응(Real-time PCR)을 이용한 융해 곡선 분석결과 KPC 25균주, NDM은 8균주에서 증폭을 확인하였고, PCR 결과와 100% 일치함을 확인하였다. 결론적으로 real-time PCR을 이용한 신속하고 특이성이 높은 CRE의조기진단은 공중보건 및 감염 중에 항균 관리접근법을 통한 병원내 감염확산 방지 및 통제가 가능할 것으로 사료된다.

Abstract

This study investigated the use of real-time PCR melting curves for the diagnosis of blaKPC and blaNDM genes among the most frequently detected carbapenemase-producing Enterobacteriaceae in Korea. As a means of addressing the shortcomings of phenotype tests and conventional PCR. The modified Hodge test confirmed positivity in 25 of 35 strains, and carbapenemase inhibition testing confirmed positivity in 14 strains by meropenem+PBA or meropenem+EDTA. PCR analysis showed amplification products in 25 strains of Klebsiella pneumoniae carbapenemases (KPC), 10 of K. pneumoniae, 5 of E. coli, 5 of A. baumannii, 4 of P. aeruginosa, and 1 of P. putida. New Delhi metallo β-lactamase (NDM) identified amplification products in 8 strains, that is, 2 K. pneumoniae, 3 E. coli, 1 P. aeruginosa, 1 E. cloacae, and 1 P. retgeri strains. Real-time PCR melting curve analysis confirmed amplification in 25 strains of KPC and 8 strains of NDM, and these results were 100% consistent with PCR results. In conclusion, our findings suggest early diagnosis of carbapenem resistant Enterobacteriaceae by real-time PCR offers a potential means of antibacterial management that can prevent and control nosocomial infection spread.

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