rep-PCR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

目的分析重复序列引物聚合酶链反应(rep-PCR)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K-B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rep-PCR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rep-PCR法可用于医院院内感染的流行病学调查。

Preliminary application of rep-PCR in epidemiological research of extended spetrum β -lactamase (ESBL) + Escherichia coli

Objective To establish better reaction factors for rep-PCR and to investigate if the extended spectrum β-lactamase(ESBL)+ Escherichia coli isolated from different patients in the respiratory ward have the same origin by using rep-PCR.Methods The ESBL confirmation was taken by double disc confirmatory test.The susceptibility testing was performed with K-B test.By applying the widespread enterobacter repetitive intergenic consensus as a primer,the stains were typed by rep-PCR following electrophoresis in agarose gel.Results The analysis of the PCR productions indicated that it would create useful DNA ban-prints and all these 22 ESBL(+)Escherichia coli strains were of three origins.Conclusion The method of rep-PCR is practical for epidemiological research in nosocomial infection.