改良Hodge试验检测肠杆菌科细菌碳青霉烯酶的性能评估  

Performance of modified Hodge test on the detection of carbapenemase among Enterobacteriaceae

作　　者：杨启文[1] 郑瑞[2] 王辉[1] 陈民钧[1] 徐英春[1] 吴伟元[3] 俞云松[4] 孙自镛[5] 童明庆[6] 张嵘[7] 廖康[8] 曹彬[9] 黄心宏[10] 朱莲娜[11] 倪语星[12] 季萍[13] 卓超[14] 苏丹虹[15] 范红[16] 刘文恩 徐修礼[18] 孙宏莉[1] 谢秀丽[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院检验科,100730 [2]云南省第一人民医院检验科 [3]深圳市人民医院实验医学科 [4]浙江大学医学院附属邵逸夫医院感染科 [5]华中科技大学同济医学院附属同济医院检验科 [6]江苏省人民医院检验科 [7]浙江大学医学院附属第二医院检验科 [8]广州中山大学附属第一医院检验科 [9]首都医科大学附属北京朝阳医院感染科 [10]福建医科大学附属协和医院检验科 [11]广西医科大学附属第一医院检验科 [12]上海交通大学附属瑞金医院临床微生物科 [13]新疆医学院附属医院检验科 [14]广州呼吸疾病研究所 [15]广州医学院第一附属医院检验科 [16]四川大学附属华西医院检验科 [17]湘雅医学院附属第一医院检验科 [18]第四军医大学附属西京医院检验科

出　　处：《中华检验医学杂志》2010年第12期1122-1127,共6页Chinese Journal of Laboratory Medicine

摘　　要：目的 评估改良Hodge试验在碳青霉烯类药物敏感性降低的肠杆菌科细菌中检测碳青霉烯酶的效能.方法 收集2004-2008年我国16家教学医院的49株碳青霉烯类药物敏感性降低（亚胺培南、美罗培由或厄他培南的MIC≥2μg/ml）的肠杆菌科细菌;采用对倍琼脂稀释法测定其对亚胺培南、美罗培南和厄他培南的MIC;通过改良Hodge试验检测碳青霉烯酶;利用加苯硼酸或苯唑西林的改良Hodge试验区分碳青霉烯酶或AmpCs/ESBLs导致的阳性结果 ;采用PCR检测包括NDM-1型碳青霉烯酶基因在内的多种β内酰胺酶基因,并对PCR阳性产物测序鉴定.结果 49株菌中,36株菌对亚胺培南不敏感（MIC〉4μg/ml）,31株菌对美罗培由不敏感（MIC〉4μg/ml）,47株菌对厄他培南不敏感（MIC〉2μg/ml）.49株临床分离菌中23株菌改良Hodge试验刚性,包括9株弱阳性和14株强阳性.经过对菌株进行β内酰胺酶基因PCR和测序,发现9株Hodge弱阳性菌株中2株携带blaKPC-2,7株不携带碳青霉烯酶基因,但携带blaampC/blaESBL;14株强阳性菌株中4株携带blaKPC-2,8株携带blaIMP-4,2株携带blaIMP-8;26株改良Hodge试验阴性菌株均未携带碳青霉烯酶基因.所有49株菌均未检测到blaNDM-1.以碳青霉烯酶基因检测为标准,则改良Hodge试验对检测碳青霉烯类药物敏感性降低的肠杆菌科菌中碳青霉烯酶的敏感度为100%,特异度为79%,阳性预测值为70%,阴性预测值为100%,准确性为86%.结论 改良Hodge试验具有很好的敏感性,但存在一些假阳性结果 .利用苯硼酸和苯唑西林可有效区分改良Hodge试验的假阳性和真阳性.Objective To evaluate the performance of modified Hodge test on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Methods Fortynine Enterobacteriaceae isolates with decreased susceptibility to carbapenems （ MIC of imipenem, meropenem or ertapenem was ≥ 2 μg/ml ） were collected from 16 teaching hospitals from 2004 to 2008. MICs of imipenem, meropenem and etapenem were determined by agar dilution method. Carbapenemases were detected by modified Hodge test. Carbepenemase-causing positive results and AmpCs-causing positive results were differentiated by phenyl boronic acid and oxacillin. Beta-lactamases encoding genes including blaNDM-1were detected by PCR and sequencing. Results Thirty-six of 49 isolates were non-susceptible to imipenem （MIC 〉4 μg/ml）, 31 were non-susceptible to meropenem （MIC 〉 4 μg/ml） and 47 were non-susceptible to ertapenem （MIC 〉 2 μg/ml）. Twenty-three isolates showed positive modified Hodge test result, including 9 weak-positive results and 14 strong-positive results. Through PCR detection and sequencing, 2 out of 9 isolates showing weak-positive results carried blaKPC-2 and other 7 did not carry any carbapenemase genes but AmpCs/ESBLs genes. Among the 14 isolates showing strong-positive results, 4 carried blaKPC-2, 8 carried blaIMP-4 and 2 caried blaIMP-8. All 26 isolates with negative modified Hodge test result didn＇t carry any carbapenemase genes. No isolate carried blaNDM-1. Carbapenemases genes PCR detection was regarded as a gold standard, and the sensitivity, specificity, positive predictive value and negative predictive value of modified hodge test was 100%, 79%, 70% and 100% on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Conclusions Modified Hodge test revealed great sensitivity but showed a few false positive results. True and false positive results can be effectively differentiated by phynel boronic acid and oxacillin.

关 键 词：肠杆菌科 Β内酰胺酶类 细菌蛋白质类 卡巴配能类 微生物敏感性试验 

分 类 号：R686[医药卫生—骨科学]