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

Kojima, Saburo; Kageyama, Tsutomu; Fukushi, Shuetsu; Hoshino, Fuminori B.; Shinohara, Michiyo; Uchida, Kenzo; Natori, Katsuro; Takeda, Naokazu; Katayama, Kazuhiko

doi:10.1016/s0166-0934(01)00404-9

Cited by 741 publications

(542 citation statements)

References 25 publications

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“…This primers have demonstrated to produce a specific amplicon after a single round of PCR (70%) reducing the detection times (data not shown). This last result disagrees with Kojima et al [2002] that report a lower detection capability of primers to target the central region of RdRp, after the first step of PCR (31%), in a broader range of NoV GII strains. The sequence analysis of VP1 region D of the genome has been obtained from three samples.…”

Section: Characterization Of the Outbreak In Ballshcontrasting

confidence: 99%

Molecular characterization of norovirus GII strains identified in Albania

Donia

Cenko²,

Divizia

2013

Journal of Medical Virology

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Noroviruses (NoVs) are considered as the leading cause of diarrheal diseases in all groups of age. In the last decade the number of NoV outbreaks worldwide is increasing. Data published by the systems of NoV surveillance show the GII.4 strain as the human predominant genotype circulating worldwide and new genetic variants of GII.4 were associated with epidemic events. In Albania the economy transformation has damaged significantly the environment and a large circulation of enteric viruses was reported in the past with the presence of NoV among the genotyped strains. This study aimed to characterize, by molecular analysis, the NoV GII strains detected in Albania during two time periods: in 2010 from the outbreak occurred in Ballsh and in 2002–2003 from sporadic cases of diarrhoea. A total of 21 Nov GII strains were characterized. The NoV GII.4 was genotyped more frequently and it was related closely to the pandemic variants recorded in GenBank. During 2002–2003, six NoV GII recombinant strains have been characterized

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Section: Characterization Of the Outbreak In Ballshcontrasting

confidence: 99%

Molecular characterization of norovirus GII strains identified in Albania

Donia

Cenko²,

Divizia

2013

Journal of Medical Virology

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“…The primer sets SR80/NVP110 [19] and JV12/JV13 [20] were used to amplify a fragment of the RNA polymerase gene of sapoviruses and noroviruses, respectively. The primer sets G1SKF/G1SKR and G2SKF/G2SKR [21] were used to amplify a fragment of the capsid gene of genogroup I and II noroviruses, respectively. The Aichi virus RT-PCR used the primers Ai6261/6779 targeting the RNA polymerase gene [22].…”

Section: Laboratory Methodsmentioning

confidence: 99%

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Lorrot

Bon

Hajje

et al. 2010

Eur J Clin Microbiol Infect Dis

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International audienceRotavirus is recognised as the most important agent of severe acute gastroenteritis (AGE) in young children. In a 2-year prospective survey, we investigated the epidemiology and clinical features of the viral and bacterial pathogens in children hospitalised for AGE. The study was performed in a Parisian teaching hospital from November 2001 to May 2004. Clinical data were prospectively collected to assess the gastroenteritis severity (20-point Vesikari severity score, the need for intravenous rehydration, duration of hospitalisation). Stools were systematically tested for group A rotavirus, norovirus, astrovirus and adenovirus 40/41, sapovirus and Aichi virus and enteropathogenic bacteria. A total of 457 children (mean age 15.9 months) were enrolled. Viruses were detected in 305 cases (66.7%) and bacteria in 31 cases (6.8%). Rotaviruses were the most frequent pathogen (48.8%), followed by noroviruses (8.3%) and adenoviruses, astroviruses, Aichi viruses and sapoviruses in 3.5%, 1.5%, 0.9% and 0.4%, respectively. Cases of rotavirus gastroenteritis were significantly more severe than those of norovirus with respect to the Vesikari score, duration of hospitalisation and the need for intravenous rehydration. Rotaviruses were the most frequent and most severe cause in children hospitalised for AGE, and noroviruses also account for a large number of cases in this population

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“…Conventional RT-PCR was carried out to sequence the real-time RT-PCR-positive specimens. Briefly, for norovirus GI PCR, primers G1SKF and G1SKR were used, and for norovirus GII PCR, primers G2SKF and G2SKR were used (17). RT-PCR products were excised from the gel and purified with the QIAquick gel extraction kit (Qiagen, Germany).…”

Section: Methodsmentioning

confidence: 99%

Norovirus Infections in Symptomatic and Asymptomatic Food Handlers in Japan

Ozawa¹,

et al. 2007

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Noroviruses are the leading cause of outbreaks of gastroenteritis in the world. At present, norovirus genogroup II, genotype 4 (GII/4), strains are the most prevalent in many countries. In this study we investigated 55 outbreaks and 35 sporadic cases of norovirus-associated gastroenteritis in food handlers in foodcatering settings between 10 November 2005 and 9 December 2006 in Japan. Stool specimens were collected from both symptomatic and asymptomatic individuals and were examined for norovirus by real-time reverse transcription-PCR; the results were then confirmed by sequence analysis. Norovirus was detected in 449 of 2,376 (19%) specimens. Four genogroup I (GI) genotypes and 12 GII genotypes, including one new GII genotype, were detected. The GII/4 sequences were predominant, accounting for 19 of 55 (35%) outbreaks and 16 of 35 (46%) sporadic cases. Our results also showed that a large number of asymptomatic food handlers were infected with norovirus GII/4 strains. Norovirus GII had a slightly higher mean viral load (1 log unit higher) than norovirus GI, i.e., 3.81 ؋ 10 8 versus 2.79 ؋ 10 7 copies/g of stool. Among norovirus GI strains, GI/4 had the highest mean viral load, whereas among GII strains, GII/4 had the highest mean viral load (2.02 ؋ 10 8 and 7.96 ؋ 10 9 copies/g of stool, respectively). Importantly, we found that asymptomatic individuals had mean viral loads similar to those of symptomatic individuals, which may account for the increased number of infections and the predominance of an asymptomatic transmission route.The positive-sense polyadenylated single-stranded RNA virus family Caliciviridae contains four genera: Norovirus, Sapovirus, Lagovirus, and Vesivirus (1). The prototype strain of human norovirus is the Norwalk virus (NV/Human/US/1968), which was first discovered in an outbreak of gastroenteritis in an elementary school in Norwalk, OH, in 1968 (15). Noroviruses are the leading cause of outbreaks of gastroenteritis in the world; they cause outbreaks in various settings, including hospitals, cruise ships, schools, and restaurants (2,9,12,15,23,24,29). In addition, noroviruses have been detected in environmental samples (e.g., treated and untreated sewage) as well as in contaminated foods such as oysters, shellfish, sandwiches, salads, raspberries, and even ice (7,18,19,26). Numerous molecular epidemiological studies have revealed a global distribution of these viruses (25,27,31).The most widely used method of detecting noroviruses is reverse transcription-PCR (RT-PCR), which has high sensitivity; also, the products can be used for further genetic analysis. Real-time RT-PCR assays have also been developed; they are sensitive, broadly reactive, and rapid for the detection of human noroviruses in clinical stool specimens and environmental samples (13,14,21).As the detection methods become more and more sensitive, the numbers of genogroups and genotypes are expected to increase. One emerging characteristic is that strains have been found to persist in one geographical region, only to disapp...

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

Cited by 741 publications

References 25 publications

Molecular characterization of norovirus GII strains identified in Albania

Molecular characterization of norovirus GII strains identified in Albania

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Norovirus Infections in Symptomatic and Asymptomatic Food Handlers in Japan

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