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카바페넴 내성 Enterobacterales에서carbapenemase의 신속 검출 및 형별 분류를 위한두 가지 상용화된 제품의 평가

Evaluation of Two Commercial Kits for Rapid Detection and Typing of Carbapenemase in Carbapenem-Resistant Enterobacterales

김미나|정윤실|허규화|성흥섭|이승후
대한임상미생물학회 (Korean Society of Clinical Microbiology)|2021.06|Annals of Clinical Microbiology|제24권 제2호|pp. 45-53|DOI 10.5145/ACM.2021.24.2.3

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GenotypePCRImmunoassayCarbapenemaseEnterobacterales

초록

배경: Carbapenemase 생성 Enterobacterales (CPE)를 신속하게 검출하는 것은 적절한 항균제 선택과 감염관리에 있어서 중요하다. KPC, NDM, VIM, IMP, OXA-48 계열 carbapenemase 검출능을 평가하기 위하여 다중 lateral flow immunoassay 기법을 사용하는 NG-Test Carba5 (Carba5; NGBiotech, France)와 전자동 실시간 PCR 기법을 사용하는 BD MAX Check-Points CPO Assay (CPO;BD Diagnostic Systems, USA) 등 2가지 검사법을 수기 PCR을 이용한 유전자형 분석과 비교평가하였다. 방법: 2013년부터 2019년까지 서울아산병원에서 분리된 카바페냄 내성Enterobacterales (CRE) 중가급적 carbapenemase 형별 당 20주 이상, carbapenemase 음성인 CRE는 60주 정도를 연구대상에포함하고자 하였다. Carbapenemase 유전자형 분석을 위해 KPC, NDM, OXA-48 계열의 경우 단독PCR을 시행하였고, VIM, IMP, GIM, SIM, SPM은 다중 PCR을 시행하였다. 모든 분리주는 Carba5와 CPO로 검사하였다. 유전자형 검사 결과가 불일치할 경우 단일 목표 유전자에 특이적인 수기PCR과 GeneXpert Carba-R Assay (Carba-R; Cepheid, USA)를추가로시행하였다. 결과: 총 147주의 CRE를 평가하였고 82주(55.8%)의 CPE를 포함하였다. CPE는 KPC 20주,NDM 22주, VIM 17주, IMP 3주, OXA-48계열 13주, 두 종류의 carbapenemase 양성인 7주(KPC/VIM 3주, NDM/VIM 2주, KPC/NDM 1주, NDM/OXA-48-계열 1주)로 구성되었다. Carba5와CPO는 모든 CPE를 정확하게 검출하였으며, 2주의 IMP 양성 CPE가 추가로 검출되었다. Carba5와 CPO의 민감도와 특이도는 둘 다 각각 100%, 97%로 동일하였다. IMP 위양성인 2주는Carba-R과 IMP-특이적인단일유전자 PCR에서 IMP 양성임을확인하였다. 결론: Carba5와 CPO는 배양된 집락으로부터 다섯 가지 주요 carbapenemase를 정확하게 검출하고형별을 분류할 수 있었다. Carba5는 더 신속하고 간편하여 임상검사실에서 즉석검사로 사용할 수있다는장점이있다.

Abstract

Background: Rapid detection of carbapenemase-producing Enterobacterales (CPE) isdesirable to guide antimicrobial therapy and infection control. The NG-Test Carba5 (Carba5;NG Biotech, France) rapid multiplex lateral flow immunoassay and BD MAX Check-Points CPOAssay (CPO; BD Diagnostic Systems, USA) fully automated real-time PCR assay were evaluatedfor the detection of KPC, NDM, VIM, IMP, and OXA-48-like group in a culture colony comparedto genotyping using conventional PCR. Methods: Among the clinical isolates of carbapenem-resistant Enterobacterales(CRE) collected from 2013 to 2019, up to 20 isolates for each carbapenemase type,and approximately 60 carbapenemase-negative CRE were enrolled. Genotyping ofcarbapenemases were performed using single-target PCR for KPC, NDM, and OXA-48-likegroup and the multiplex PCR for VIM, IMP, GIM, SIM, and SPM. All isolates were tested withCarba5 and CPO. The discrepant results were resolved by single-target specific conventionalPCR or GeneXpert Carba-R Assay (Carba-R; Cepheid, USA). Results: Of 147 CREs, 82 were CPE (55.8%) including 20 KPC, 22 NDM, 17 VIM, three IMP, and13 OXA-48-like group, and seven double carbapenemase-positive (three KPC/VIM, two NDM/VIM, one KPC/NDM, and one NDM/OXA-48-like group) isolates. Carba5 and CPO detected allCPE correctly along with two more IMP-producing CPE. The sensitivity and specificity of bothkits were equally 100% and 97%. Two false IMP-positives were confirmed IMP-positive withCarba-R and IMP-specific single-target PCR. Conclusion: Carba5 and CPO reliably detect and differentiate five common carbapenemasesin cultured colonies. Carba5, faster and simpler, is preferred as a spot test.

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