产超广谱β-内酰胺酶细菌微量表型确证试验及其应用评价

Establishment of a Rapid Micromethod for Phenotypic Confirmatory Test of Extended-Spectrum β-Lactamases Producing Bacteria

目的建立产超广谱β-内酰胺酶(ESBL)确证的快速微量检测法,评价该方法在筛检工作中的应用效果。方法以灭菌的96孔酶标板建立ESBL微量快速确证试验,用已知59株产ESBL大肠埃希菌、33株非产ESBL大肠埃希菌和质控菌株进行试验的真实性、可靠性和收益评价。结果重复试验符合率100%;用酶标仪自动判读敏感度达到或超过94.92%,特异度和阳性预测值均为100%,与美国国家实验室标准化研究所(CLSI)推荐标准检测方法所得到的结果差别无统计学意义(P>0.05)。结论ESBL微量确证试验具有良好的真实性、可靠性和检测效益;操作简便、经济、快捷。

Objective To establish a rapid micromethod for phenotypic confirmatory test of extended-spectrum β-lactamases （ESBL） producing bacteria and to evaluate the efficacy of the method in screening programme. Methods The advanced rapid micromethod for the confirmation of ESBL producing strains was designed using sterile 96 well plastic plates based on the principal of ESBL phenotypic confirmatory test recommended by the current Clinical and Laboratory Standards Institute（CLSI）. The testretest reliability and validity, as well as the effectiveness of the micromethod were assessed by the randomized controlled trial consisting of 59 ESBL positive E. coli, 33 ESBL negative E. coli, and 2 quality control strains. Results Sufficiently high test-retest reliability and validity was achieved with a coincident rate of 100%, a specificity and positive predictive value of 100%, and a sufficient sensitivity beyond 94.92% by the automatic enzyme-labeled assay at an optical density of 570, 600 or 630 nm. Conclusion An extraordinary efficiency for the rapid micromethod of phenotypic confirmatory test of ESBL was established.