3-氨基苯硼酸联合乙二胺四乙酸碳青霉烯酶抑制增强试验检测肠杆菌目细菌产碳青霉烯酶的研究  

Detection of carbapenemase production in Enterobacterales by carbapenemase inhibitor enhancement test using phenylboronic acid and EDTA

作　　者：周银娣 郭燕[1] 张琴 韩仁如 胡付品 ZHOU Yindi;GUO Yan;ZHANG Qin;HAN Renru;HU Fupin(Institute of Antibiotics,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所,上海200040 [2]合肥市第一人民医院检验科 [3]成都市妇女儿童中心医院院感科

出　　处：《中国感染与化疗杂志》2022年第1期65-71,共7页Chinese Journal of Infection and Chemotherapy

摘　　要：目的了解3-氨基苯硼酸(PBA)联合乙二胺四乙酸(EDTA)碳青霉烯酶抑制剂增强试验(PBA-EDTA法)用于检测肠杆菌目细菌产碳青霉烯酶的检测结果。方法使用PBA-EDTA法测定275株经金标准PCR及DNA测序已明确为产碳青霉烯酶肠杆菌目细菌产碳青霉烯酶结果的符合率,并与CLSI推荐的mCIM联合eCIM改良碳青霉烯灭活试验(mCIMeCIM法)检测结果的符合率比较。结果PBA-EDTA法对产KPC酶菌株的符合率为99.2%(118/119),对产金属酶菌株的符合率为100%(109/109),对产KPC+MβL复合酶的符合率为100%(13/13);但不能检出34株D组OXA酶;其检测结果与分子生物学检测结果总体符合率为87.3%(240/275)。mCIM-eCIM法检测结果显示产KPC酶菌株符合率100%(119/119);检测金属酶的符合率94.5%(103/109);未能检出13株产KPC+MβL复合酶,虽能检出34株产D组OXA酶菌株产酶特性但不能分类酶型;与分子生物学检测结果相比较,mCIM-eCIM法检测结果总体符合率为93.1%(256/275)。两种方法对于KPC酶和金属酶的检出虽有差异,但显示此差异无显著统计学意义。PBA-EDTA法对复合酶(KPC+金属酶)的检出优于mCIM-eCIM法。结论PBA-EDTA法可以检测细菌产生的A组丝氨酸碳青霉烯酶、B组金属酶以及同时产A组和B组碳青霉烯酶的菌株;操作较mCIM-eCIM法简单易行;便于临床微生物实验室使用和开展肠杆菌目细菌产生的丝氨酸碳青霉烯酶和金属酶的检测,为临床合理使用抗菌药物、精准治疗患者提供实验室依据。Objective This study aimed to evaluate the reliability of carbapenemase inhibitor enhancement test using phenylboronic acid (PBA) and EDTA (PBA-EDTA assay) for detection of carbapenemases in Enterobacterales.Methods This method was used to determine carbapenemase production in 275 strains of carbapenemase-producing Enterobacterales strains,which were confirmed by gold standard PCR and DNA sequencing.The results were compared with the results of modified carbapenem inactivation test (mCIM) combined with EDTA-modified carbapenem inactivation test (eCIM) (mCIM-eCIM assay) recommended by the Clinical and Laboratory Standards Institute.Results PBA-EDTA assay accurately detected 99.2% (118/119) of the KPC-producing strains,100% (109/109) of the metallo-β-lactamase-producing strains,and 100% (13/13) of the strains producing both KPC type and metallo-β-lactamase,but did not identify the class D (OXA-type) β-lactamases in 34 strains.The PBA-EDTA assay achieved an overall coincidence rate of 87.3% (240/275) compared with the results of molecular biology test in detection of carbapenemase production.The mCIM-eCIM assay identified 100% (119/119) of the KPC-producing strains and 94.5% (103/109) of the metallo-β-lactamase-producing strains,but did not identify the 13 strains producing both KPC type and metallo-β-lactamase.The mCIM-eCIM assay detected the 34 strains producing β-lactamases,but did not identify the class D (OXA type) enzyme.The mCIM-eCIM assay resulted in an overall coincidence rate of 93.1% (256/275) compared with the results of molecular biology test in detection of carbapenemase production.The two assays performed slightly and insignificantly different in detection of KPC type and metallo-β-lactamases.The PBA-EDTA assay was better than the mCIM-eCIM method in identifying the strains producing both KPC type and metallo-β-lactamases.Conclusions The PBA-EDTA assay is useful for detection of the strains producing class A (serine carbapenemase),and/or class B (metallo-β-lactamases) β-lactamases.It is easi

关 键 词：碳青霉烯酶 碳青霉烯类耐药肠杆菌目细菌 碳青霉烯酶抑制增强试验 

分 类 号：R446.5[医药卫生—诊断学]