Jiang X, Wang J, Estes M K
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas, USA.
Arch Virol. 1995;140(2):363-74. doi: 10.1007/BF01309870.
Stool samples from 451 patients involved in volunteer studies, 26 outbreaks and approximately 175 sporadic cases of acute gastroenteritis from different geographical locations in the world were tested for Norwalk virus (NV) using a newly developed antigen ELISA and RT-PCR. NV was detected in most outbreaks previously characterized as being of NV origin. Overall, a low number of positives for NV was obtained using either RT-PCR with primers that amplified a unique region of the genome, or an ELISA with hyperimmune antisera made to the baculovirus-expressed recombinant NV capsid. However, a significant number of positives was obtained when these samples were tested by RT-PCR using primers that amplified the more highly conserved regions of the genome. Sequence analysis of the amplified viral cDNAs indicated that small round structured viruses (SRSVs) with a wide range of variable genomic sequences (44-87% nucleotide and 31-99% amino acid similarity to the 8Flla NV genome sequence) were responsible for these outbreaks. Several recent outbreaks from the US, Japan and the UK were related to the Snow Mountain Agent (SMA) by sequence analyses. Continued accumulation of sequence information will facilitate the design of new primers for virus detection and increase our understanding of the relationships and epidemiology of these viruses from different sources.
使用新开发的抗原酶联免疫吸附测定法（ELISA）和逆转录-聚合酶链反应（RT-PCR），对来自参与志愿者研究的451名患者、26起疫情以及世界各地约175例散发性急性胃肠炎病例的粪便样本进行了诺沃克病毒（NV）检测。在大多数先前被认定为NV起源的疫情中检测到了NV。总体而言，使用能扩增基因组独特区域的引物进行RT-PCR，或使用针对杆状病毒表达的重组NV衣壳制备的超免疫抗血清进行ELISA，获得的NV阳性数量都很低。然而，当使用能扩增基因组中保守性更高区域的引物通过RT-PCR对这些样本进行检测时，获得了大量阳性结果。对扩增的病毒cDNA进行的序列分析表明，具有广泛可变基因组序列（与8Flla NV基因组序列的核苷酸相似性为44%-87%，氨基酸相似性为31%-99%）的小圆结构病毒（SRSV）是这些疫情的病原体。通过序列分析，美国、日本和英国近期的几起疫情与雪山因子（SMA）有关。序列信息的持续积累将有助于设计用于病毒检测的新引物，并增进我们对这些来自不同来源的病毒之间的关系和流行病学的理解。
