Clancy Leighton E, Craig Maria E, White Peter A, Rawlinson William D
Virology Division, Department of Microbiology, SEALS, The Prince of Wales Hospital, Randwick, Sydney, NSW, Australia.
J Med Virol. 2005 Jun;76(2):215-22. doi: 10.1002/jmv.20344.
Genotyping and serotyping of enteroviruses is important for epidemiological, prognostic, and therapeutic reasons. In this study clinical isolates of enterovirus 71 during an outbreak of childhood meningoencephalitis in Sydney, Australia were identified using heteroduplex mobility analysis (HMA) of products from RT-PCR amplification of the 5' untranslated region. Five enterovirus 71 isolates shared identical heteroduplex patterns and nucleotide sequences in the 5' untranslated region. A sixth isolate exhibited minor differences in heteroduplex pattern and sequencing confirmed the isolate varied by 1% at the nucleotide level. The use of multiple reference strains and the analysis of heteroduplex patterns increased the confidence of isolate identification, and allowed identification of strain variation which could be subsequently further analyzed using sequencing. HMA can be used to accurately distinguish identical and variant isolates derived from sporadic cases and clustered infections with enteroviruses, including those causing serious infections.
出于流行病学、预后和治疗方面的原因,肠道病毒的基因分型和血清分型很重要。在本研究中,利用对5'非翻译区RT-PCR扩增产物进行异源双链迁移分析(HMA),对澳大利亚悉尼儿童脑膜脑炎暴发期间的肠道病毒71型临床分离株进行了鉴定。5株肠道病毒71型分离株在5'非翻译区具有相同的异源双链模式和核苷酸序列。第6株分离株的异源双链模式存在微小差异,测序证实该分离株在核苷酸水平上有1%的差异。使用多个参考菌株以及对异源双链模式的分析提高了分离株鉴定的可信度,并能够鉴定出菌株变异,随后可使用测序对其进行进一步分析。HMA可用于准确区分散发病例和肠道病毒聚集性感染(包括那些引起严重感染的病例)中产生的相同和变异分离株。