IRS-PCR在分析不动杆菌DNA异质性中的应用

Application of in Infrequent-Restriction-Site PCR Analysising the DNA Heterogeneity of Clinical Isolates of Acinetobacter

目的采用低频限制性位点聚合酶链反应技术(IRS-PCR)对20株临床分离的不动杆菌DNA进行异质性分析。方法通过PCR扩增稀有限制区附近DNA序列,对20株临床分离的不动杆菌进行基因分型,并与RAPD方法的分型及生化表型结果进行比较分析。结果20株不动杆菌因生化性状不同分为溶血不动杆菌(10株),洛菲不动杆菌(7株)及其他(3株);IRS-PCR将该溶血不动杆菌分为5个基因型,洛菲不动杆菌分为4个基因型,其他不动杆菌分为2个基因型;RAPD方法,引物1扩增结果将溶血不动杆菌分为6个基因型,洛菲不动杆菌分为4个基因型,其他不动杆菌分为2个基因型;引物2扩增结果将溶血不动杆菌分为9个基因型,洛菲不动杆菌分为5个基因型,其他不动杆菌分为2个基因型。结论IRS-PCR可用于不动杆菌DNA异质性的分析,且较生化表型精细、准确,与RAPD技术比较两者分辨率相当,但IRS-PCR具有较好的稳定性和重复性,值得临床推广。

Objective To analyse the DNA heterogeneity of 20 clinical isolates of Acinetobacteria using infrequet - restriction - site PCR （ IRS - PCR ）. Methods Strain - specific electrophoretic patterns from PCR products by amplifying DNA sequences flanking infre-quent restriction of 20 strains of Acinetobacter were compared with the results of genotype with RAPD as well as the results of biotyping. Results Among the 20 bacteria, ten can be recognized as Acinetobacter haemolytius with biotyping, seven as Acinetobacter lwoffi, while the other 3 bacteria can not be recognized. Acinetobacter haemolytius isolates were divided into 5 gene types with IRS- PCR, isolates of Acinetobacter lwoffi were divided into 4 gene tvoes, meanwhile, the last three bactria were divided into 2 gene types. With RAPD tech-nique, Acinetobacter haemolytius, Acinetobacter lwoffi and the other 3 bacteria were divided into 6 gene types, 4 types, and 2 types by prime 1, respectively, the amplifying results of primer 2 divided Acinetobacter haemolytius into 9 gene types, the Acinetobacter lwoffi ,5 types and the other 3 bacteria,2 types. Conclusion IRS-PCR can be used for typing Acinetobacter and is more aceurate than biotyping. It has the same recogniton ability as RAPD while is of better stability and repeatability, so this method is capable of clinical application.