Genogroup-specific PCR primers for detection of Norwalk-like viruses

被引:688
作者
Kojima, S
Kageyama, T
Fukushi, S
Hoshino, FB
Shinohara, M
Uchida, K
Natori, K
Takeda, N
Katayama, K
机构
[1] Natl Inst Infect Dis, Dept Virus Dis & Vaccine Control, Tokyo 2080011, Japan
[2] BML, R&D Ctr, Infect Dis Sect, Kawagoe, Saitama 3501101, Japan
[3] Saitama Inst Publ Hlth, Urawa, Saitama 3380824, Japan
[4] Natl Inst Infect Dis, Dept Virol 2, Tokyo 1628640, Japan
关键词
NLV; RT-PCR; genogroup-specific detection; high-sensitivity detection;
D O I
10.1016/S0166-0934(01)00404-9
中图分类号
Q5 [生物化学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
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.
引用
收藏
页码:107 / 114
页数:8
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