刘景武 王亚倩 王超 种克 张蕊

【摘 要】目的：建立一種简便的可直接用于临床尿液标本中，由碳青霉烯耐药肠杆菌科细菌引起尿路感染的主动筛查方法，并对耐药基因进行分型。方法：使用建立的方法对人工模拟的样本进行检测，确定其检测灵敏度。对临床样本采用培养结合PCR的方法和免疫层析方法分别进行检测，以培养结合PCR的方法作为金标准，评价免疫层析方法作为直接检测临床尿液标本中碳青霉烯耐药肠杆菌耐药基因快速分型的性能。结果：采用免疫层析方法检测人工模拟样本中的KPC酶和NDM酶，其中携带KPC酶的肺炎克雷伯氏菌检测灵敏度为1×104CFU/ml， 携带NDM酶的大肠埃希氏菌检测灵敏度为1×104CFU/ml。检测了100份临床尿液标本，与培养结合PCR方法相比较，采用免疫层析法直接检测临床尿液标本灵敏度为100%，特异性为100%，阳性预测值为100%，阴性预测值为100%，Youden，s指数为1。结论：采用离心结合免疫层析方法，可直接用于尿标本中碳青霉烯耐药基因的筛查和基因类型。

【关键词】碳青霉烯酶，分型，PCR，免疫层析

A simple and convenient method for active screening of urinary tract infection caused by carbapenem-resistant enterobacteriaceae bacteria

Liu Jingwu1， Wang Yaqian1， Wang Chao2， Zhong Ke3， Zhang rui1 1.Department of clinical laboratory， the third hospital of shijiazhuang， Hebei Shijiazhuang 050001， China；2. Department of Laboratory Medicine， Shijiazhuang People’s Hospital， Hebei Shijiazhuang 050001， China；3.Shijiazhuang Vocational College of Technology Information ， Hebei Shijiazhuang 050001， China

【Abstract】Objective：To establish a simple and direct screening method for urinary tract infection caused by carbapenemse-resistant enterobacteriaceae in clinical urine specimens， and to type the drug-resistant genes. Methods：The established method was used to detect the artificial simulated samples and determine their detection sensitivity. The culture-bound PCR method and the immunochromatography method were used to detect the clinical samples respectively.The culture-bound PCR method was used as the gold standard.The analysis software was used to analyze the detection results of the immunochromatography and evaluate its performance as a direct method to detect the rapid typing of carbapenemse-resistant enterobacteriaceae. Results：KPC enzyme and NDM enzyme in artificial simulated samples were detected by immunochromatography. Klebsiella pneumoniae carrying KPC enzyme was detected at a sensitivity of 1×104CFU/ mL， and Escherichia coli carrying NDM enzyme was detected at a sensitivity of 1×104CFU/ mL. 100 clinical urine specimens were tested， compared with the culture combined with PCR method，the sensitivity and specificity of immunochromatography were 100%， positive predictive value was 100%， negative predictive value was 100%， and Youden， S index was 1.Conclusion： The centrifugation combined with immunochromatography can be directly used to screen clinical urine samples for the presence of enterobacteriaceae bacteria carrying carbapenem resistance genes as well as the types of drug-resistant genes.

【Key？Words】Carbapenemase； Typing；PCR； Immunochromatography

由耐碳青霉烯类肠杆菌科细菌（CRE）造成的感染呈现逐年上升的趋势。目前的检测方法，不能很好地满足临床快速筛查的需求。免疫层析方法具有操作简便，检测速度快等特点，本研究采用免疫层析方法产品通过建立一套简单的前处理方法直接用于临床尿液标本中碳青霉烯耐药肠杆菌科细菌耐药基因判断，并评价其应用价值[1-2]。

1.1 菌株来源

菌株分离自我院住院病人标本。……