Genogroup-specific PCR primers for detection of Norwalk-like viruses
Genogroup-specific PCR primers for detection of Norwalk-like viruses
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DOI：
10.1016/s0166-0934(01)00404-9
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发表时间：
2002-02-01
影响因子：
3.1
通讯作者：
Katayama, K
中科院分区：
文献类型：
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作者：
Kojima, S;Kageyama, T;Katayama, K
诺瓦克样病毒（NLV）是非细菌性胃肠炎的主要病原体。目前仍有许多NLV菌株难以通过普通的逆转录-聚合酶链反应进行基因扩增。这主要是由于NLV之间的遗传多样性，特别是引物序列的不匹配，这限制了该技术的临床应用。本研究开发了基于衣壳区域的改进引物集，以基因组特异性的方式检测基因组I和基因组II NLV。用该引物对电镜下NLV阳性的胃肠炎患者粪便标本进行检测，所有标本RT-PCR均为阳性。前述引物对RdRp和衣壳蛋白的扩增率分别为31%和77%。因此，新的引物对NLV引起的非细菌性肠胃炎的扩增和快速诊断以及流行病学研究非常有用。(C) 2002 Elsevier Science B.V.版权所有
Norwalk-like viruses (NLV) are a major causative agent of nonbacterial gastroenteritis. There are still many NLV strains that are refractory to gene amplification by ordinary reverse transcription-polymerase chain reaction. This is due mainly to the genetic diversity among NLV, especially mismatches in the primer sequences, which limits this technique in clinical utility. In this study, improved primer sets based on the capsid region, to detect both genogroup I and II NLV by genogroup-specific manner, were developed. When stool specimens from gastroenteritis patients, that were positive for NLV by electron microscopy, were tested by this new primer set, all specimens were positive by RT-PCR. Primers described previously for RdRp and capsid protein were capable of amplifying the specimens by 31 and 77%, respectively. Therefore, new primer sets are extremely useful for the amplification and rapid diagnosis of nonbacterial gastroenteritis due to NLV as well as for epidemiological studies. (C) 2002 Elsevier Science B.V. All rights reserved.