repPCR法在检测产超广谱β内酰胺酶大肠艾希氏菌院内流行中的应用
大肠艾希氏菌,,大肠艾希氏菌;聚合酶链反应;流行病学,1材料与方法,2结果,3讨论,参考文献
摘要:目的 分析重复序列引物聚合酶链反应(repPCR)的优化条件,并采用本方法分析本院呼吸科病房不同患者痰标本中分离到的产超广谱β内酰胺酶(ESBL)大肠杆菌是否起源于同一菌株。方法 用双纸片确定法测定是否产ESBL;用KB法测定对抗生素的敏感程度;选用肠道菌广泛存在的基因间重复片断为引物进行扩增,并对实验条件进行优化;对产ESBL大肠艾希氏菌进行基因分型分析。结果 建立了repPCR法方法,显示可将大肠艾希氏菌扩增出丰富的区带。分离到的22株产ESBL大肠艾希氏菌分属3个基因型。结论 repPCR法可用于医院院内感染的流行病学调查。关键词:大肠艾希氏菌;聚合酶链反应;流行病学
Preliminary application of repPCR in epidemiological research of extended spetrum βlactamase (ESBL)+ Escherichia coli
Ma Lieting, Yang Duo, Wang Yawen
(Department of Clinical Laboratory, The First Hospital of Xian Jiaotong University, Xian 710061, China)
ABSTRACT: Objective To establish better reaction factors for repPCR and to investigate if the extended spectrum βlactamase (ESBL)+ Escherichia coli isolated from different patients in the respiratory ward have the same origin by using repPCR. Methods The ESBL confirmation was taken by double disc confirmatory test. The susceptibility testing was performed with KB test. By applying the widespread enterobacter repetitive intergenic consensus as a primer, the stains were typed by repPCR following electrophoresis in agarose gel. Results The analysis of the PCR productions indicated that it would create useful DNA banprints and all these 22 ESBL(+)Escherichia coli strains were of three origins. Conclusion The method of repPCR is practical for epidemiological research in nosocomial infection. ......
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