IRS-PCR在鲍曼不动杆菌基因分型研究中的应用

探讨低频限制性位点聚合酶链反应(infrequent-restriction-site polym erase chain site,IRS-PCR)在鲍曼不动杆菌基因分型中的应用。用IRS-PCR分别扩增上海、哈尔滨两地各20株鲍曼不动杆菌DNA稀有限制区旁序列并分型。与RAPD法比较两种基因分型方法的分型率、分辨力、重复性。IRS-PCR可将上海株分13个型别,RAPD分为19个型别,两种方法分型的一致率为60%(12/20);IRS-PCR将哈尔滨株分成19个型别,RAPD分为20个型别,两种方法分型的一致率为90%(18/20)。与RAPD相比,IRS-PCR的条带数多,分辨力强,重复性好。故IRS-PCR可用于鲍曼不动杆菌的分型研究,也是一种有价值的分子流行病学研究工具。

Application of Infrequent-restriction-site PCR as a Typing Method of Acinetobacter baumannii

This study is to explore the value of infrequent-restriction-site PCR(IRS-PCR) as a typing method of Aninetobacter baumannii.By using IRS-PCR,the sequences of flanking infrequent restriction sites in the DNAs from 40 Acinetobacter baumannii isolates(20 strains collected from Harbin,the rest collected from Shanghai) were amplified and genotyped.The typing and discriminatory abilities as well as reproducibility of IRS-PCR detection were compared with those of RAPD.The 20 isolates from shanghai could be divided into 13 genotypes by IRS-PCR and 19 genotypes by RAPD and the accordance ratio was 60%(12/20).The other 20 isolates from Harbin could be classified into 19 genotypes by IRS-PCR and 20 genotypes by RAPD and the accordance ratio was 90%(18/20).In comparison with RAPD,IRS-PCR offered more amplification fragments,stronger discrimination and higher reproducibility.As a conclusion,IRS-PCR is suitable for genotyping Aninetobacter baumannii.Furthermore,IRS-PCR can be used as a valuable tool for molecular epidemiological research.