Jordens J Z, Leaves N I, Anderson E C, Slack M P
Public Health Laboratory Service Haemophilus Reference Laboratory, University of Oxford, United Kingdom.
J Clin Microbiol. 1993 Nov;31(11):2981-7. doi: 10.1128/jcm.31.11.2981-2987.1993.
A polymerase chain reaction-based typing method for noncapsulate Haemophilus influenzae was developed. Randomly amplified polymorphic DNA fingerprints were generated from boiled supernatants prepared directly from bacterial colonies without the need for DNA extraction. The technique was applied to isolates obtained during putative outbreaks of chest infection and validated by comparison with sodium dodecyl sulfatepolyacrylamide gel electrophoresis analysis of outer membrane protein-enriched preparations and rRNA gene restriction analysis. There was complete concordance between the three techniques. The results show that randomly amplified polymorphic DNA analysis provides a highly discriminatory method of characterizing strains of noncapsulate H. influenzae which is eminently suitable as an epidemiological tool for the rapid investigation of outbreaks of infection.
开发了一种基于聚合酶链反应的非包膜型流感嗜血杆菌分型方法。直接从细菌菌落制备的煮沸上清液中生成随机扩增多态性DNA指纹,无需进行DNA提取。该技术应用于在假定的胸部感染暴发期间获得的分离株,并通过与富含外膜蛋白制剂的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析和rRNA基因限制性分析进行比较来验证。这三种技术之间完全一致。结果表明,随机扩增多态性DNA分析提供了一种高度区分性的方法来鉴定非包膜型流感嗜血杆菌菌株,非常适合作为快速调查感染暴发的流行病学工具。