【作者】 王沛； 何永贵；

【Author】 WANG Pei HE Yonggui(The First People’s Hospital of Jingmen,Hubei 448000,China)

【机构】 湖北省荆门市第一人民医院；

【摘要】 目的建立荧光原位杂交法,快速鉴定临床标本中的粪肠球菌和屎肠球菌。方法将荧光同位素标记的分别针对粪肠球菌和屎肠球菌的核糖体16srRNA及23srRNA序列互补的寡核苷酸探针,在同一张玻片上杂交30min至1h,用杂交缓冲液洗涤10min,干燥后置显微镜下观察。结果探针的特异性经标准菌株证实,对163份临床标本测试结果与培养法结果比较,两种探针的特异性和敏感性均为100%。结论荧光原位杂交法适宜于临床标本中粪肠球菌和屎肠球菌的快速鉴定。更多还原

【Abstract】 OBJECTIVE To establish a new protocol for rapid identification of E.faecalis and E.faecium in clinical samples by Fluorescent In Situ Hybridization.METHODS By using oligonucleotide probes,fluorescently labeled with fluorescein isothiocyanate,targets the 16srRNA or 23srRNA of E.faecalis and E.faecium respectively,the mixed probes were added to one slide and hybridized about 30 min to one hour at 50℃,washed unbound probes away with washing buffer about 10min at 50℃,slide was observed under fluoresceince microscope. RESULTS The specificity of two probes was validated by reference strains.163 strains from clinical isolates were tested,the method showed 100%specificity and sesitivity for E.faecalis and E.faecium.CONCLUSIONS The result indicates FISH allow rapid identification of E.faecalis and E.faecium in clinical samples.更多还原