MALDI-TOF MS直接靶板微滴生长法对耐碳青霉烯类肠杆菌目细菌的诊断价值  

Value of MALDI-TOF MS direct-on-target growth assay in diagnosis of carbapenem-resistant Enterobacteriaceae

作　　者：黄声雷 曹锦楠 周春妹 马艳 潘柏申 王蓓丽 郭玮 胡必杰 HUANG Shenglei;CAO Jinnan;ZHOU Chunmei;MA Yan;PAN Baisheng;WANG Beili;GUO Wei;HU Bijie(Zhongshan Hospital,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学附属中山医院检验科,上海200032 [2]复旦大学附属中山医院感染病科,上海200032

出　　处：《中华医院感染学杂志》2025年第6期812-817,共6页Chinese Journal of Nosocomiology

基　　金：病原微生物生物安全国家重点实验室开放基金资助项目(SKLPBS2144);白求恩基金中青年医生优才培养计划基金资助项目(BCF-SY-0311-20190807-07)。

摘　　要：目的评估基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)直接鉴定阳性血培养瓶中耐碳青霉烯类大肠埃希菌和肺炎克雷伯菌的价值。方法收集2021年7月-2023年6月复旦大学附属中山医院血流感染患者中大肠埃希菌或肺炎克雷伯菌阳性的血培养瓶,使用分离胶管收集菌体,将受试菌分别与4μg/ml亚胺培南、4μg/ml美罗培南和2μg/ml厄他培南溶液共培养,35℃孵育4和5 h后,再用质谱鉴定。使用微量肉汤稀释法检测三种药物的最低抑菌浓度,并作为本试验金标准。结果共收集31株大肠埃希菌和28株肺炎克雷伯菌阳性的血培养瓶,其中大肠埃希菌4和5 h孵育后的对照生长有效率均为100.00%,肺炎克雷伯菌4和5 h孵育后的对照生长有效率分别为96.43%和100.00%。5 h孵育后检测肺炎克雷伯菌对三种药物耐药的灵敏度、特异性、阳性和阴性预测值均为100.00%。5 h孵育后检测大肠埃希菌对亚胺培南、美罗培南和厄他培南耐药的特异性及阳性预测值均为100.00%,灵敏度分别为73.58%、78.93%和78.93%,阴性预测值分别为70.60%、75.00%和75.00%。结论使用质谱直接鉴别阳性血培养中耐碳青霉烯类大肠埃希菌和肺炎克雷伯菌结果快速准确,尤其5 h孵育时间后检测耐药的肺炎克雷伯菌准确性100.00%,该方法为临床及时治疗由耐碳青霉烯类肠杆菌目细菌引起的血流感染提供依据。OBJECTIVE To evaluate the value of matrix-assisted laser desorption ionization time-of-flight-mass spectrometry(MALDI-TOF MS)in direct identification of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae positive for blood culture.METHODS The blood culture bottles that were positive for E.coli or K.pneumoniae were collected from the patients with bloodstream infection who were treated in Zhongshan Hospital,Fudan University from Jul.2021 to Jun.2023.The isolates were collected by using a gel-contact clotting tube,then the tested strains were respectively mixed with 4μg/ml of imipenem,4μg/ml of meropenem and 2μg/ml of ertapenem for coculture and were incubated at 35℃for 4 and 5 hours,finally,the strains were identified by using the mass spectrum.The minimum inhibitory concentrations(MICs)of the three types of drugs were tested by microbroth dilution method and were set as the golden standards for the test.RESULTS Totally 31 strains of E.coli and 28 strains of K.pneumoniae that were positive in blood culture bottles were collected,both of the effective rates of controlled growth of the E.coli strains were 100.00%after the incubation for 4 and 5 hours,and the effective rates of controlled growth of the K.pneumoniae strains were 96.43%and 100.00%after the incubation for 4 and hours,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of the K.pneumoniae strains against the three types of drugs were 100.00%after the incubation for 5 hours.The specificity and positive predictive value of the E.coli strains against imipenem,meropenem and ertapenem were 100.00%after the incubation for 5 hours;the sensitivities were 73.58%,78.93%and 78.93%,respectively;the negative predictive values were 70.60%,75.00%and 75.00%,respectively.CONCLUSIONS MALDI-TOF MS is rapid and accurate for direct identification of the carbapenem-resistant E.coli and K.pneumoniae strains positive in blood culture bottles,and the accuracy reaches at 100.00%for the test of drug-resistant K.pneumon

关 键 词：基质辅助激光解析电离飞行时间质谱 直接靶板微滴生长法 阳性血培养 直接鉴定 耐碳青霉烯类肠杆菌科细菌 

分 类 号：R446.5[医药卫生—诊断学]