Detection of hepatitis A virus in shellfish by nested reverse transcription-PCR

Croci, Luciana; Medici, Dario De; Morace, G; Fiore, Alfonsina; Scalfaro, Concetta; Beneduce, Francesca; Toti, L.

doi:10.1016/s0168-1605(99)00028-8

Cited by 49 publications

(30 citation statements)

References 19 publications

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“…Moreover, preliminary steps, such as concentration of viruses from the sample and nucleic acid purification (removal of inhibitors), are essential for final PCR accuracy and reproducibility. Different methods have been proposed for determining viral contamination based on whole shellfish (8,29) or dissected tissue (4,50) for all types of virus (4,22,24) or for specific viruses (9,10,27). The method based on dissected tissues (4) is considered to be specific, reliable, and reproducible (3), providing a nucleic acid extract that allows detection of most enteric viruses (33).…”

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confidence: 99%

Three-Year Study To Assess Human Enteric Viruses in Shellfish

Guyader

Haugarreau

Miossec

et al. 2000

Appl Environ Microbiol

252

134

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The main pathogenic enteric viruses able to persist in the environment, such as hepatitis A virus (HAV), Norwalk-like virus (NLV), enterovirus (EV), rotavirus (RV), and astrovirus (AV), were detected by reverse transcription-PCR and hybridization in shellfish during a 3-year study. Oyster samples (n ‫؍‬ 108), occasionally containing bacteria, were less frequently contaminated, showing positivity for AV (17%), NLV (23%), EV (19%), and RV (27%), whereas mussel samples, collected in areas routinely impacted by human sewage, were more highly contaminated: AV (50%), HAV (13%), NLV (35%), EV (45%), and RV (52%). Sequences obtained from HAV and NLV amplicons showed a great variety of strains, especially for NLV (strains close to Mexico, Snow Mountain Agent, or Norwalk virus). Viral contamination was mainly observed during winter months, although there were some seasonal differences among the viruses. This first study of virus detection over a fairly long period of time suggests that routine analysis of shellfish by a molecular technique is feasible.

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mentioning

confidence: 99%

Three-Year Study To Assess Human Enteric Viruses in Shellfish

Guyader

Haugarreau

Miossec

et al. 2000

Appl Environ Microbiol

252

134

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“…Both for viral suspensions and experimentally contaminated shellfish, RNA extraction was performed with Nucleospin RNAII (Macherey-Nagel, Düren, Germany) according to manufacturer instructions, and nucleic acid was eluted in a 100-l volume. For naturally contaminated samples 300 l of shellfish extract, equivalent to 5 g of shellfish homogenate, was extracted as described by Afzal and Minor (1) and Croci et al (7). The RNA pellet was resuspended in 10 l of deionized triply distilled water containing 1 U of RNase inhibitor/l.…”

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confidence: 99%

Reverse Transcription-Booster PCR for Detection of Noroviruses in Shellfish

Medici

Croci

Suffredini

et al. 2004

Appl Environ Microbiol

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The methods commonly used for norovirus (NV) detection are based on reverse transcription-PCR (RT-PCR) followed by confirmation of the amplified sequence. To increase sensitivity, an RT-booster PCR was developed. The proposed method showed an increase in sensitivity at least 2 log units for all the NV strains tested compared with the standard RT-PCR method. Higher sensitivity was confirmed in tests on experimentally and naturally contaminated shellfish.

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“…Shellfish consumption in Korea is a major risk factor for HAV infection [31], since these products are commonly eaten raw or slightly cooked. Only a drastic heat treatment can assure a complete inactivation of virus [32] and only through cooking of shellfish can reduce the risk to human health [25]. This makes clear the potential to public concern about shellfish safety.…”

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confidence: 99%

Monitoring the Hepatitis A Virus in Oyster from Korea

Park¹,

Jeong²,

Jung³

et al. 2015

AiM

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Hepatitis A virus (HAV) causes many cases of oyster-or clam-associated gastroenteritis in various countries. HAV was detected on oyster by RT-PCR in 19.6% (11/56) in Korea. The percentages of HAV-positive samples in 2011 and 2012 were 27.6% and 11.1%, respectively. Phylogenetic analysis revealed that several nucleotide sequences highly similar to those of HAVs isolated in this study. Phylogenetic analysis of the coding regions of the viral protein VP4/VP2 revealed that all amplicons were classified into IA genogroup. It will provide useful data that aids in our understanding circulating HAVs and may contribute to future control.

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Detection of hepatitis A virus in shellfish by nested reverse transcription-PCR

Cited by 49 publications

References 19 publications

Three-Year Study To Assess Human Enteric Viruses in Shellfish

Three-Year Study To Assess Human Enteric Viruses in Shellfish

Reverse Transcription-Booster PCR for Detection of Noroviruses in Shellfish

Monitoring the Hepatitis A Virus in Oyster from Korea

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