Detection and Sequencing of Norwalk‐Like Viruses from Stool Samples in Japan Using Reverse Transcription‐Polymerase Chain Reaction Amplification

Nakayama, Miyuki; Ueda, Yuhichi; Kawamoto, Hiroyoshi; Han-jun, Yue; Saito, Kunihikó; Nishio, Osamu; Ushijima, Hiroshi

doi:10.1111/j.1348-0421.1996.tb03343.x

Cited by 16 publications

(11 citation statements)

References 21 publications

(38 reference statements)

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“…On the basis of sequence analysis of the 81-bp RNA polymerase amplicon, Chc168-09/97, Art189-01/98, and Shr241-02/ 98 were grouped in genogroup I, as described elsewhere [4,25,26]. However, the analysis presented in this report from a larger polymerase region (figure 3) and by others from capsid sequence analyses [11] supports the classification of Chc168-09/97 as genogroup II.…”

Section: Discussionsupporting

confidence: 70%

Genetic Diversity and Molecular Epidemiology of Norwalk‐Like Viruses

2000

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Specimens (n=287) from 59 gastroenteritis outbreaks collected from February 1997 to March 1999 were analyzed by reverse transcriptase-polymerase chain reaction. The majority of outbreaks (88%) were associated with Norwalk-like viruses. Molecular analyses of strains from 46 outbreaks showed the cocirculation during the 1998-1999 winter of 2 genogroup II clusters, accounting for 57% and 28% of outbreaks, respectively. An important genetic diversity was observed during this 2-year period. Thirteen different genogroup II strains and 3 different genogroup I strains were found. Genogroup I strains, although from the same cluster, were highly divergent (9%-16%). Epidemiologic and molecular data indicate that several introductions did not result in any major shift of prominent strains, whereas 1 apparently established itself. Some point mutations allowed corroboration of epidemiologic links and strongly suggest that, in several instances, sharing staff and/or transfer of patients between health care institutions can create a significant risk for Norwalk-like virus dissemination.

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Section: Discussionsupporting

confidence: 70%

Genetic Diversity and Molecular Epidemiology of Norwalk‐Like Viruses

2000

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“…Stool samples were determined previously to be bacteria-, rotavirus-, and enteric adenovirus-negative by the latex agglutination test and enzyme immunoassay (EIA) [Ushijima et al, 1992;Ushijima et al, 1994]. Calicivirus was screened by RT-PCR with calicivirusspecific primers [Nakayama et al, 1996].…”

Section: Stool Samplesmentioning

confidence: 99%

Molecular epidemiology of astroviruses in Japan from 1995 to 1998 by reverse transcription-polymerase chain reaction with serotype-specific primers (1 to 8)

et al. 2000

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In addition to the serotype-specific primers described previously (1 to 7), a new serotype 8-specific primer has been designed, allowing detection of all astrovirus serotypes. A total of 1,382 diarrheal stool samples in 5 regions in Japan were examined by reverse transcription-polymerase chain reaction (RT-PCR). The incidence of astrovirus infection in all 5 regions was 5.9% (82 of 1,382 samples) and infection occurred mainly from November to April. Serotypes 1, 3, and 4 were detected in 66, 14, and 2 of the 82 positive samples, respectively. None of the other serotypes was detected. The highest detection rate was from 0 to 1 year old, 39.0%, and the next highest was from 1 to 2 years old, 34.1%. The primers provide a useful approach for study of the epidemiology of astroviruses.

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“…The great sequence diversity of the NLV genome has made this a challenging undertaking. Primers for previous assays have been taken from open reading frame 1 (ORF1) of the RNA-dependent RNA polymerase (RdRp) gene (1,2,9,13,15,26,27,31,37) or from ORF2 of the capsid protein gene (6,10,11,13,28,38,39).…”

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confidence: 99%

Broadly Reactive and Highly Sensitive Assay for Norwalk-Like Viruses Based on Real-Time Quantitative Reverse Transcription-PCR

Kageyama

Kojima

Shinohara³

et al. 2003

J Clin Microbiol

1,296

889

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We have developed an assay for the detection of Norwalk-like viruses (NLVs) based on reverse transcription-PCR (RT-PCR) that is highly sensitive to a broad range of NLVs. We isolated virus from 71 NLV-positive stool specimens from 37 outbreaks of nonbacterial acute gastroenteritis and sequenced the open reading frame 1 (ORF1)-ORF2 junction region, the most conserved region of the NLV genome. The data were subjected to multiple-sequence alignment analysis and similarity plot analysis. We used the most conserved sequences that react with diverse NLVs to design primers and TaqMan probes for the respective genogroups of NLV, GI and GII, for use in a real-time quantitative RT-PCR assay. Our method detected NLV in 99% (80 of 81) of the stool specimens that were positive by electron microscopy, a better detection rate than with the two available RT-PCR methods. Furthermore, our new method also detected NLV in 20 of 28 stool specimens from the same NLV-related outbreaks that were negative for virus by electron microscopy. Our new assay is free from carryover DNA contamination and detects low copy numbers of NLV RNA. It can be used as a routine assay for diagnosis as well as for elucidation of the epidemiology of NLV infections.

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Detection and Sequencing of Norwalk‐Like Viruses from Stool Samples in Japan Using Reverse Transcription‐Polymerase Chain Reaction Amplification

Cited by 16 publications

References 21 publications

Genetic Diversity and Molecular Epidemiology of Norwalk‐Like Viruses

Genetic Diversity and Molecular Epidemiology of Norwalk‐Like Viruses

Molecular epidemiology of astroviruses in Japan from 1995 to 1998 by reverse transcription-polymerase chain reaction with serotype-specific primers (1 to 8)

Broadly Reactive and Highly Sensitive Assay for Norwalk-Like Viruses Based on Real-Time Quantitative Reverse Transcription-PCR

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