mCIM和eCIM联合检测产碳青霉烯酶肠杆菌科细菌的应用

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目的:探讨改良碳青霉烯灭活试验(modified carbapenem inactivation method, mCIM)联合EDTA改良碳青霉烯灭活试验(EDTA-modified carbapenem inactivation method, eCIM)检测感染患者分离的耐碳青霉烯肠杆菌科细菌 (Carbapenem-resistant n Enterobacteriaceae, CRE)碳青霉烯酶的方法对诊控CRE感染的应用价值。n 方法:回顾性分析2017年1至12月天津医科大学总医院临床感染患者分离的78株非重复对碳青霉烯类抗生素耐药的肠杆菌科细菌的耐药情况,应用Vitek2 Compact全自动细菌鉴定仪鉴定至种并检测其对厄他培南和亚胺培南的最低抑菌浓度(minimum inhibitory concentration, MIC),用分子生物学PCR试验检测其碳青霉烯酶基因n blaKPC、n blaNDM、n blaIMP和n blaVIM,基因测序确定基因型作为金标准,同时采用mCIM联合eCIM试验对所收集的细菌进行碳青霉烯酶检测。以PCR结果为标准,计算mCIM和eCIM的灵敏度、特异度、阳性预测值、阴性预测值及Kappa值,进行mCIM和eCIM联合检测结果与PCR结果的一致性检验。n 结果:78株碳青霉烯耐药肠杆菌科细菌应用PCR法检测出碳青霉烯酶基因阳性74株,阴性4株,其中n blaKPC-2阳性60株,n blaNDM-1阳性2株,n blaNDM-5阳性7株,n blaNDM-9阳性3株,n blaIMP-4阳性1株,n blaKPC-2和 n blaNDM-1同时阳性1株。mCIM检测碳青霉烯酶的敏感度为100%,特异度为100%, Kappa值为1.000;mCIM和eCIM联合检测丝氨酸碳青霉烯酶敏感度为100%,特异度为100%,Kappa值为1.000;联合检测金属碳青霉烯酶敏感度为92.9%,特异度为100%, Kappa值为0.955。n 结论:采用mCIM、eCIM联合试验对CRE中的碳青霉烯酶进行检测,实用性强,结果易于判读,依据CRE细菌中基因型的不同,将为临床CRE诊断与抗菌药物精准治疗和感染控制提供重要诊断依据。“,”Objective:To explore the clinical application value of modified carbapenem inactivation test (mCIM) combined with EDTA-modified carbapenem inactivation test (eCIM) for detecting the carbapenemase of CRE isolated from infected patients in clinical diagnosis and infection control of CRE infection.Methods:Drug resistance of seventy eight non-repetitive enterobacteriaceae resistant to carbapenem antibiotics, which were isolated from clinically infected patients from January 2017 to December 2017 in the Tianjin Medical University General Hospital, was retrospectively analyzed. Meanwhile, Vitek2 Compact automatic bacterial identification instrument was used to identify the species and detect its minimum inhibitory concentration (MIC) for ertapenem and imipenem. Carbapenemase genes n blaKPC, n blaNDM, n blaIMP and n blaVIM were detected by PCR test, the genotype was determined by gene sequencing as the gold standard, and mCIM Combined eCIM test was used for carbapenemase detection of collected bacteria. Using PCR results as the standard, the sensitivity, specificity, positive predictive value, negative predictive value and Cohen′ Kappa value of mCIM and eCIM were calculated, and the consistency of the combined detection results of mCIM and eCIM and PCR results was checked.n Results:Seventy eight carbapenem-resistant Enterobacteriaceae were detected by PCR, 74 carbapenemase gene positive and 4 negative, including 60 n blaKPC-2 positive, 2 n blaNDM-1 positive and n blaNDM-5 positive Of the 7 strains, 3 strains were positive for n blaNDM-9, 1 strain was positive for n blaIMP-4, and 1 strain was both positive for n blaKPC-2 and n blaNDM-1. The sensitivity of mCIM to detect carbapenemase is 100%, the specificity is 100%, and the Kappa value is 1.000; the combined detection of mCIM and eCIM for serine carbapenemase has a sensitivity of 100%, the specificity is 100%, and the Kappa value is 1.000; The sensitivity of combined detection of metallo-β-lactamase is 92.9%, the specificity is 100%, and the Kappa value is 0.955.n Conclusions:The combined test of mCIM and eCIM which is used to detect carbapenemase in CRE costs low, doesn′t require special reagents and equipment, has strong practicability, simple operation, and easy interpretation of results. According to the different genotypes of CRE bacteria, it provides important clinical diagnostic evidence for clinical CRE diagnosis, precise antimicrobial treatment and infection control.
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