金黄色葡萄球菌随机扩增DNA多态性分型研究

Typing of Staphylococcus aureus by random amplification of polymorphic DNA

目的建立金黄色葡萄球菌随机扩增ＤＮＡ多态性（ＲＡＰＤ）分型技术，以应用于临床菌种鉴定及流行病学调查。方法对临床分离的１５株金黄色葡萄球菌，酚氯仿法提取其ＤＮＡ后行ＲＡＰＤ分型，同时进行噬菌体分型。结果１５株临床分离株大多数产生独特而稳定的ＲＡＰＤ带型，可分成５个噬菌体型和１４个ＲＡＰＤ谱型。采用同一反应体系，产肠毒素金黄色葡萄球菌未出现ＲＡＰＤ谱型。结论ＲＡＰＤ基因分型技术不需已知核酸序列，分型率高，分辨力强，简便快捷，可为金黄色葡萄球菌临床分离株提供分型标志，是分子流行病学研究的有效方法。

OBJECTIVE To establish the random amplification of ploymorphic DNA (RAPD) typing method of S.aureus and to apply it in the clinical strain identification and epidemiology investigation were studied. METHODS DNA from 15 clinical strains of S.aureus were extracted by phenol-chloroform method and used in the RAPD assay, phage typing were also performed. RESULTS Almost all the clinical strains showed stable and distinct RAPD pattern and were divided into 5 phagetypes and 14 RAPD profiles, while enterotoxin-producing S.aureus failed to show RAPD pattern using this assay. CONCLUSION RAPD provides markers for typing of clinical strains and is suitable for molecular epidemiologic studies with high typeability, powerful discrimination, simplicity and rapidness.