乳胶法、胶体金法和实时荧光定量PCR法在6种碳青霉烯酶检测中的价值及评价

姜懿桐; 高春艳

doi:10.3784/jbjc.202411140655

乳胶法、胶体金法和实时荧光定量PCR法在6种碳青霉烯酶检测中的价值及评价

Performance of latex method, colloidal gold immunochromatography and quantitative real-time polymerase chain reaction in detection of six carbapenemases

摘要

摘要:
目的评价乳胶法、胶体金法和实时荧光定量聚合酶链式反应（qPCR）法在6种常见碳青霉烯酶检测中的性能及价值，为临床和公共卫生实验室进行碳青霉烯酶快速检测方法的选择提供可靠依据。
方法对收集的236株菌株同时使用乳胶法、胶体金法和qPCR法进行6种常见碳青霉烯酶检测，评价3种方法的一致性。
结果 3种方法检测肺炎克雷伯菌碳青霉烯酶（KPC）均阳性菌株58株（24.58%）；1株菌KPC检测不一致，胶体金法检测结果为阴性，乳胶法和qPCR法为阳性。3种方法检测新德里金属-β-内酰胺酶（NDM）均阳性菌株37株（15.68%），维罗纳整合素编码金属β-内酰胺酶（VIM）均阳性菌株10株（4.24%），亚胺培南酶（IMP）均阳性菌株15株（6.36%），苯唑西林酶23（OXA23）均阳性菌株45株（19.06%），苯唑西林酶48（OXA48）均阳性菌株10株（4.24%）。用乳胶法和胶体金法检测OXA23有4株与qPCR结果不一致，OXA48有2株与qPCR结果不一致，不一致的结果均为qPCR结果检出相应碳青霉烯酶基因，而乳胶法和胶体金法检测结果为阴性。 3种方法检测指示同一菌株携带多种酶的有11株菌，其中，KPC和NDM均为阳性的有3株；NDM和IMP均为阳性的有1株；KPC和OXA48均为阳性的有1株；NDM和OXA48均为阳性的有2株；VIM和IMP均为阳性的有4株。
结论碳青霉烯酶乳胶法、胶体金法检测和qPCR法比较，3种方法一致性好。乳胶法因不需要额外仪器且检测速度快，适合基层单位检测碳青霉烯酶，具有广阔的应用前景。值得关注的是同时携带多种碳青霉烯酶菌株的检出，以防超级耐药菌在临床的传播和流行。

Abstract:
Objective To systematically evaluate the performance of latex method, colloidal gold immunochromatography and quantitative real-time polymerase chain reaction (qPCR) in the detection of six carbapenemases, and provide reliable evidence for the selection of rapid detection assays for carbapenemase in clinical and public health laboratories.
Methods A total of 236 strains were used for six carbapenemases detection by latex method, colloidal gold immunochromatography and qPCR and the consistency of the three methods was evaluated.
Results In this study, the detection results with 3 methods indicated that 58 strains were positive for Klebsiella pneumoniae carbapenemase (KPC) (24.58%), and 1 strain had inconsistent KPC detection results, which was negative by colloidal gold immunochromatography and positive by latex method and qPCR, 37 strains were positive for New Delhi metallo-β-lactamase (NDM) (15.68%), 10 strains were positive for Verona integron-encoded metallo-β-lactamase (VIM)(4.24%), 15 strains were positive for imipenemase (IMP) (6.36%), 45 strains were positive for oxacillinases 23(OXA23)(19.06%) and 10 strains were positive for oxacillinases 48 (OXA48)(4.24%). Four strains had inconsistent detection results of OXA23 by latex method and colloidal gold immunochromatography (negative) and qPCR (positive), and 2 strains had inconsistent detection results of OXA48 by latex method and colloidal gold immunochromatography (negative) and qPCR (positive). Additionally, there were 11 strains which were multiple enzyme positive by three methods, in which 3 were positive for both KPC and NDM, 1 was positive for both NDM and IMP, 1 was positive for both KPC and OXA48, 2 were positive for both NDM and OXA48 , and 4 were positive for both VIM and IMP.
Conclusion There were good consistency among the three detection methods. As a new carbapenemase genotyping detection method, latex method is simple and easy to use in the simultaneous and rapid detections of KPC, OXA48, OXA23, VIM, IMP and NDM, which can facilitate clinical diagnosis and treatment, and it is necessary to pay attention to the detection of multiple enzymes for the prevention of the spread of drug resistance.

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