NORWALK-LIKE VIRUSES - DEMONSTRATION OF GENOMIC DIVERSITY BY POLYMERASE CHAIN-REACTION
NORWALK-LIKE VIRUSES - DEMONSTRATION OF GENOMIC DIVERSITY BY POLYMERASE CHAIN-REACTION
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DOI：
10.1128/jcm.31.11.3007-3012.1993
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发表时间：
1993-11-01
影响因子：
9.4
通讯作者：
BROWN, DWG
中科院分区：
文献类型：
--
作者：
GREEN, J;NORCOTT, JP;BROWN, DWG
建立了一种检测粪便中诺瓦克样病毒的逆转录-聚合酶链反应（RT-PCR）方法。对在英国收集的99份粪便标本进行了检测，这些粪便标本通过电子显微镜检查确定含有小的圆形结构病毒颗粒。它们来自50起爆发和16起散发的病毒性胃肠炎病例。在三次暴发的15份标本中检测到诺瓦克病毒RNA大小合适的RT-PCR产物，表明与诺瓦克病毒密切相关的病毒近年来在英国没有广泛传播。对4个分离株的两个基因组区域的RT-PCR扩增产物进行测序，比较基因组变异程度。PCR产物的DNA测序显示，来自英国的毒株之间存在很强的相似性（扩增的两个区域约为97%），但与诺瓦克病毒存在显著差异（67%至78%）。所有RT-PCR检测的病毒均经固相免疫电镜或酶联免疫吸附试验鉴定为血清型UK 2。这些发现为诺瓦克病毒和血清型UK 2小圆结构病毒之间的基因组关系提供了证据。
A reverse transcription-polymerase chain reaction (RT-PCR) amplification procedure was developed for the detection of Norwalk-like viruses in fecal specimens. Ninety-nine fecal specimens collected in the United Kingdom and containing small round-structured virus particles as determined by electron microscopy were tested. They came from 50 outbreaks and 16 sporadic cases of viral gastroenteritis. RT-PCR products of the appropriate size for Norwalk virus RNA were detected in 15 specimens from three outbreaks, suggesting that viruses closely related to Norwalk virus have not been circulating widely in the United Kingdom in recent years. From four isolates, the RT-PCR amplification products of two genomic regions were sequenced and the degree of genomic variation was compared. DNA sequencing of the PCR products revealed strong similarities among strains from the United Kingdom (approximately 97% for both regions amplified) but significant differences from Norwalk virus (67 to 78%). All of the viruses detected by RT-PCR were classified as serotype UK2 by solid-phase immune electron microscopy or enzyme-linked immunosorbent assay. These findings provide evidence of a genomic relationship between Norwalk virus and serotype UK2 small round-structured viruses.