Virucidal effect of UV light on hepatitis a virus in sea water: evaluation with cell culture and RT-PCR

Virucidal effect of UV light on hepatitis A virus was investigated in artificial sea water. Infectious virus was no longer detectable after 15 min irradiation of 3 liter experimentally contaminated water. Genomic amplification by polymerase chain reaction after reverse transcription allowed the detection of viral RNA in all samples even after 60 min irradiation.

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Virucidal effect of UV light on hepatitis a virus in sea water: evaluation with cell culture and RT-PCR

Water Science & Technology ◽

10.1016/0273-1223(95)00257-n ◽

1995 ◽

Vol 31 (5-6) ◽

Cited By ~ 1

Keyword(s):

Cell Culture ◽

Hepatitis A ◽

Sea Water ◽

Hepatitis A Virus ◽

Uv Light ◽

Rt Pcr ◽

Virucidal Effect

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Persistence of Hepatitis A Virus in Oysters†

Journal of Food Protection ◽

10.4315/0362-028x-66.2.331 ◽

2003 ◽

Vol 66 (2) ◽

pp. 331-334 ◽

Cited By ~ 62

Author(s):

DAVID H. KINGSLEY ◽

GARY P. RICHARDS

Keyword(s):

Polymerase Chain Reaction ◽

Crassostrea Virginica ◽

Reverse Transcription ◽

Hepatitis A ◽

Infectious Virus ◽

Hepatitis A Virus ◽

Complete Removal ◽

Chain Reaction ◽

Daily Feeding ◽

Polymerase Chain

We investigated the ability of hepatitis A virus (HAV) to persist for up to 6 weeks in Eastern oysters (Crassostrea virginica). Viral RNA was detected by reverse transcription–polymerase chain reaction 6 weeks after 16 h of exposure to 90,000 PFU (180 PFU/ml of seawater) of HAV. Assaying for infectious virus in oysters that received a daily feeding of phytoplankton recovered 3,800, 650, and 500 PFU of HAV 1, 2, and 3 weeks after contamination with 90,000 PFU of HAV, respectively. However, no infectious HAV was isolated from oysters 4, 5, or 6 weeks after contamination. These results support the position that shellfish depuration is insufficient for the complete removal of infectious viruses. Extended relay times (in excess of 4 weeks) may be required to produce virologically safe shellfish.

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Hepatitis A Virus Detection in Oysters (Crassostrea gigas) in Santa Catarina State, Brazil, by Reverse Transcription–Polymerase Chain Reaction

Journal of Food Protection ◽

10.4315/0362-028x-66.3.507 ◽

2003 ◽

Vol 66 (3) ◽

pp. 507-511 ◽

Cited By ~ 29

Author(s):

C. COELHO ◽

A. P. HEINERT ◽

C. M. O. SIMÕES ◽

C. R. M. BARARDI

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Severe Illness ◽

Rt Pcr ◽

Chain Reaction ◽

Santa Catarina ◽

Polymerase Chain ◽

Santa Catarina State

Shellfish are readily contaminated with viruses present in water containing sewage because of the concentration effect of filter feeding. Hepatitis A virus (HAV) is the main cause of acute hepatitis worldwide and may lead to severe illness or even death. It is transmitted through fecal and oral routes and causes widespread endemic and asymptomatic infections in young children. Here we describe a method for the detection of HAV RNA in shellfish involving the extraction of total RNA from oyster meat followed by reverse transcription–polymerase chain reaction (RT-PCR). Virus recovery from oyster extracts artificially seeded with HAV strain HM 175 was examined by RT-PCR. The minimum detection limit was 3.3 focus-forming units of HAV, and the recovery rate was 75.7%. This method was used to assess the viral contamination of four shellfish beds in Santa Catarina State, Brazil, over a 1-year period. Six (22%) of 27 samples collected in autumn and winter from one shellfish bed tested positive for HAV.

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Development of PCR Methods for Enteric Virus Detection in Water

Water Science & Technology ◽

10.2166/wst.1993.0348 ◽

1993 ◽

Vol 27 (3-4) ◽

pp. 211-218 ◽

Cited By ~ 17

Author(s):

K. J. Schwab ◽

R. De Leon ◽

M. D. Sobsey

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Detection System ◽

Virus Detection ◽

Enteric Virus ◽

Rt Pcr ◽

Chain Reaction ◽

Sample Concentration ◽

Spin Column ◽

Polymerase Chain

This study developed a reverse transcriptase-polymerase chain reaction (RT-PCR) detection system for enteric viruses in sample concentrates obtained by conventional filter adsorption-elution methods. One liter beef extract (BE)-glycine (G) eluant seeded with poliovirus 1 and hepatitis A virus (HAV) was used as a model system and the eluant further processed for RT-PCR compatibility. Sample concentration and purification procedures consisted of polyethylene glycol (PEG) precipitation, Pro-Cipitate (Affinity Technology, New Brunswick, NJ) precipitation, a second PEG precipitation, spin-column chromatography, and ultrafiltration. Sample volumes are reduced from 1 L to 20-40 µL and purified sufficiently for viral detection by RT-PCR. As little as 3 PFU of poliovirus 1 in an initial 1 L eluate were detected by RT-PCR.

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Development of a PCR-based method for the detection of enteroviruses and hepatitis a virus in molluscan shellfish and its application to polluted field samples1

Water Science & Technology ◽

10.2166/wst.1995.0659 ◽

1995 ◽

Vol 31 (5-6) ◽

pp. 457-464 ◽

Cited By ~ 7

Author(s):

David N. Lees ◽

Kathleen Henshilwood ◽

Sarah Butcher

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Pcr Amplification ◽

Field Site ◽

Rt Pcr ◽

Pcr Inhibition ◽

Field Samples ◽

Polymerase Chain ◽

Low Levels ◽

Bivalve Shellfish

The use of the polymerase chain reaction (PCR) for detection of low levels of enteric viruses in bivalve shellfish is hindered by the presence of potent amplification inhibitors. A procedure previously developed for removing the majority of these amplification inhibitors is applied to the detection of enteroviruses and hepatitis A virus in naturally polluted field samples. Quantification of PCR inhibition showed that PCR sample tolerance ranged from 2 to 4.7g shellfish for highly polluted samples. These results indicate the need for adequate controls for PCR inhibition, particularly for negative samples. Reverse transcription (RT)-PCR results were compared with conventional enterovirus isolation for a range of naturally contaminated shellfish. All enterovirus isolation positive samples were also positive by enterovirus RT-PCR. At one field site shellfish were positive by enterovirus RT-PCR but negative for virus isolation. All shellfish tested were negative for hepatitis A by RT-PCR. The procedure for removal of PCR amplification inhibitors should be equally applicable to the detection of Norwalk and related Small Round Structured Viruses (SRSVs) in shellfish.

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ICC/PCR detection of enteroviruses and hepatitis A virus in environmental samples

Canadian Journal of Microbiology ◽

10.1139/w00-134 ◽

2001 ◽

Vol 47 (2) ◽

pp. 153-157 ◽

Cited By ~ 46

Author(s):

Kelly A Reynolds ◽

Charles P Gerba ◽

Morteza Abbaszadegan ◽

Ian L Pepper

Keyword(s):

Cell Culture ◽

Environmental Samples ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Virus Detection ◽

Rt Pcr ◽

Specific Detection ◽

Polymerase Chain ◽

Conventional Cell ◽

Polymerase Chain Reaction Methodology

This study applied the integrated cell culture/polymerase chain reaction methodology (ICC/PCR) for rapid and specific detection of both cytopathogenic and noncytopathogenic viruses. Results of this study showed that the use of direct RT-PCR or conventional cell culture alone may yield erroneous results with the analysis of environmental samples. The purpose of this study was to compare cultural, molecular, and combined assays for the most effective method of virus detection in variable environmental samples. Using ICC/PCR, stock enterovirus inocula of [Formula: see text]10 PFU were PCR positive in at least 4/5 replicate flasks after only 5 h of incubation in cell culture, and in all flasks after [Formula: see text]10 h. An inoculum of one PFU was detected by PCR after 20 h of cell culture incubation while for concentrations of virus below one PFU, 25 h of incubation was sufficient. Similarly, hepatitis A virus (HAV) inocula of 100 MPN/flask, produced indeterminate CPE in cell culture, but were clearly detected by ICC/PCR following 48 h of incubation. Lower levels of HAV, 1 and 10 MPN, were detected by ICC/PCR after 96 to 72 h of incubation, respectively. Cell culture lysates from 11 environmental sample concentrates of sewage, marine water, and surface drinking water sources, were positive for enteroviruses by ICC/PCR compared to 3 positive by direct RT-PCR alone. Results from ICC/PCR eventually agreed with cell culture but required [Formula: see text]48 h of incubation, compared to as long as 3 weeks for CPE following incubation with BGM and FRhK cells.Key words: RT-PCR, cell culture, ICC/PCR, enterovirus, hepatitis A virus.

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An outbreak of hepatitis A among homosexuals linked to a family outbreak

Epidemiology and Infection ◽

10.1017/s0950268802007069 ◽

2002 ◽

Vol 129 (1) ◽

pp. 113-117 ◽

Cited By ~ 22

Author(s):

K. STENE-JOHANSEN ◽

P. A. JENUM ◽

T. HOEL ◽

H. BLYSTAD ◽

H. SUNDE ◽

...

Keyword(s):

Molecular Epidemiology ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Sequence Information ◽

Risk Of Infection ◽

Rt Pcr ◽

Chain Reaction ◽

The Family ◽

Polymerase Chain ◽

Family Outbreak

Several outbreaks of hepatitis A occurred in Norway in 1995–8. Molecular epidemiology was used to follow the spread of hepatitis A virus in the population. Distinct strains of hepatitis A virus (HAV) were detected by reverse transcriptase–polymerase chain reaction (RT–PCR) and subsequent sequencing in serum from patients in different communities at risk of infection. Two HAV strains were detected in an outbreak among 26 men having sexual contact with other men. One of these strains was also detected in a geographically limited family outbreak. The family outbreak was first believed to be acquired abroad. The sequence information linked the two outbreaks, and epidemiological and serological analyses revealed the transmission route. This study demonstrates the importance of molecular epidemiology in outbreak investigation, surveillance and monitoring of hepatitis A in the population.

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