Detection of hepatitis A virus, rotavirus, and enterovirus in naturally contaminated shellfish and sediment by reverse transcription-seminested PCR

Guyader, F. Le; Dubois, Éric; Ménard, D.; Pommepuy, Monique

doi:10.1128/aem.60.10.3665-3671.1994

Cited by 176 publications

(61 citation statements)

References 46 publications

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“…According to our data, RTBCR is not substantially more sensitive for detecting rotavirus than other techniques such as the ELBA test used in our study, in agreement with the results obtained by Gouvea et al (1990) and Ushijima et al (1992). The use of a double amplification system, nested or semi-nested PCR, has been reported by some authors to improve the sensitivity of rotavirus detection (Gouvea et al, 1990 ;Gentsch et al, 1992 ;Le Guyader et al, 1994), though others did not observe an increase in sensitivity using these procedures (Ushijima et az., 1992).…”

Section: Discussionsupporting

confidence: 90%

“…as from environmental samples (Le Guyader et al, 1994). Our results suggest that both CFll purification and the RNAID procedure for extracting viral RNA may be used prior to performing RTKR.…”

Section: Discussionmentioning

confidence: 88%

“…Such studies will lead to a better understanding of the pathogenesis of rotavirus infections and may confirm the detection of rotavirus antigens in respiratory secretions reported by others (Fragosa et al, 1986;Zheng et al, 1991). In addition, RT/PCR is also a powerful technique to further investigate the mechanisms of transmission of these viruses and their presence in the environment (Wilde et ul., 1992;Le Guyader et d., 1994). EM continues to be a mainstay in the diagnosis of rotaviral diseases and has been used in the past as the final arbiter when discrepancies occurred with other techniques (Kapikian and Chanock, 1996).…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rota viruses: applications of the assay

Buesa

Colomina

Raga

et al. 1996

Research in Virology

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Our aim was to evaluate the reverse transcription and polymerase chain reaction (RT/PCR) technique for the detection of rotavirus shedding by infected children as a routine diagnostic procedure, in comparison to the enzyme-linked immunosorbent assay (ELISA), electron microscopy (EM) and polyacrylamide gel electrophoresis (PAGE) of rotavirus double-stranded RNA. Two-hundred and twenty stool specimens were collected from infants and young children with diarrhoea, and 10-20% faecal suspensions were made. Several methods of rotavirus dsRNA extraction were assayed. Electrophoretic analysis of viral RNA was carried out on 10% polyacrylamide gels followed by silver staining. RT/PCR was performed using oligonucleotide primers specific for both 3' and 5' ends of the rotavirus gene encoding VP7 which are highly conserved among group A rotaviruses. Following RNA extraction with phenol-chloroform and ethanol precipitation, RT/PCR could detect rotaviral RNA in only 11 of 25 samples known to contain rotaviruses by conventional methods. The purification of RNA extracts by CF11 cellulose and the application of the RNAID method were equally effective in extracting RNA and/or removing inhibitory substances from the faecal samples. RT/PCR led to the detection of 66 positive samples from 220 specimens tested (30%), whilst 64 specimens were positive by ELISA (29%), 59 (26.8%) by PAGE and 56 (25.4%) by EM. In our study, RT/PCR was 100 times more sensitive than the ELISA test in detecting rotaviruses serially diluted in a faecal suspension. Although RT/PCR is theoretically much more sensitive than ELISA, PAGE and EM for detection of rotaviruses, great care must be taken to remove inhibitory substances from the enzymatic reactions. We do not consider that RT/PCR should replace immunoassays with high sensitivity and specificity for rotavirus testing in faecal samples, although this technique has other applications, like the search for rotavirus in different clinical specimens (sera, cerebrospinal fluid, respiratory secretions, etc.) and in environmental samples, as well as the typing of viral strains using serotype-specific primers.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 99%

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Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rota viruses: applications of the assay

Buesa

Colomina

Raga

et al. 1996

Research in Virology

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“…This European directive establishes that microbial quality of shell¢sh should be monitored by measuring counts of total fecal coliform bacteria, Escherichia coli, and Salmonellae. The Directive establishes no speci¢c microbiological criteria concerning the presence of enteric viruses, even though it has clearly been shown that there is no correlation between the presence of viruses and the presence of coliform bacteria and/or E. coli; in fact HAV, enteroviruses and NLV have been detected in mussels that otherwise meet bacteriological standards [187^189, 199,200]. Similarly, it is not clear if and how depuration will reduce the levels of viral contaminants, especially for NLV, since quantitative methods for their detection are not yet available.…”

Section: Legislation Rules and Regulationsmentioning

confidence: 99%

Foodborne viruses

Koopmans¹

2002

FEMS Microbiology Reviews

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Foodborne and waterborne viral infections are increasingly recognized as causes of illness in humans. This increase is partly explained by changes in food processing and consumption patterns that lead to the worldwide availability of high-risk food. As a result, vast outbreaks may occur due to contamination of food by a single foodhandler or at a single source. Although there are numerous fecal-orally transmitted viruses, most reports of foodborne transmission describe infections with Norwalk-like caliciviruses (NLV) and hepatitis A virus (HAV), suggesting that these viruses are associated with the greatest risk of foodborne transmission. NLV and HAV can be transmitted from person to person, or indirectly via food, water, or fomites contaminated with virus-containing feces or vomit. People can be infected without showing symptoms. The high frequency of secondary cases of NLV illness and - to a lesser extent - of hepatitis A following a foodborne outbreak results in amplification of the problem. The burden of illness is highest in the elderly, and therefore is likely to increase due to the aging population. For HAV, the burden of illness may increase following hygienic control measures, due to a decreasing population of naturally immune individuals and a concurrent increase in the population at risk. Recent advances in the research of NLV and HAV have led to the development of molecular methods which can be used for molecular tracing of virus strains. These methods can be and have been used for the detection of common source outbreaks. While traditionally certain foods have been implicated in virus outbreaks, it is clear that almost any food item can be involved, provided it has been handled by an infected person. There are no established methods for detection of viruses in foods other than shellfish. Little information is available on disinfection and preventive measures specifically for these viruses. Studies addressing this issue are hampered by the lack of culture systems. As currently available routine monitoring systems exclusively focus on bacterial pathogens, efforts should be made to combine epidemiological and virological information for a combined laboratory-based rapid detection system for foodborne viruses. With better surveillance, including typing information, outbreaks of foodborne infections could be reported faster to prevent further spread.

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“…Kopecka et al 1993;Wyn-Jones et al 1995), ground waters , sludge-amended ®eld soils (Straub et al 1995), and shell®sh (e.g. Le Guyader et al 1994). The technique has been extended to cover other virus groups present in water, including adenoviruses (Puig et al 1994), HAV (Graff et al 1993), astrovirus (Marx et al 1995) and rotavirus (Gajardo et al 1995).…”

Section: Detection By Molecular Biologymentioning

confidence: 99%