Concentration of Hepatitis a Virus in Environmental Samples

1991 ◽  
Vol 24 (2) ◽  
pp. 229-234 ◽  
Author(s):  
A. Bosch ◽  
R. Gajardo ◽  
F. X. Abad ◽  
J. M. Diez ◽  
J. Jofre
Keyword(s):  
Environmental Samples ◽  
Hepatitis A ◽  
Glass Powder ◽  
Hepatitis A Virus ◽  
Tap Water ◽  
Plaque Assay ◽  
Wild Type ◽  
Filter Aid ◽  
Treatment Concentration ◽  
Positively Charged

The cytopathogenic pHM-175 strain of hepatitis A virus was used to develop different procedures for the concentration of HAV in tap water, fresh water, seawater and raw sewage, HAV was quantified by a plaque assay in the FRhK-4 cell line. Water samples were concentrated by a modification of the adsorption to and elution from glass powder (GPAE) method, by adsorption to and elution from filter aid, and by ammonium sulfate flocculation (ASF). The GPAE method consistently yielded greater HAV recoveries than filtration through filter aid, or ASF. HAV was concentrated by GPAE from 20-litre samples with satisfactory efficiencies in all kinds of water: 100% for tap water, 80% for freshwater, 75% for seawater and 61% for sewage. Concentration efficiencies for filter aid and ASF were always lower than 25% and 40%, respectively, in any kind of water. The charge of glass powder was modified by polyethylenimine treatment. Concentration efficiencies of HAV in 20 1 samples through adsorption to and elution from positively charged glass powder (PGPAE) were 100% for tap water, 94% for seawater, and 61% for freshwater and sewage. The presence of wild-type HAV in sewage samples could be monitored by molecular hybridization with cDNA probes after GPAE concentration.

Hepatitis a Virus Concentration from Experimentally Contaminated Distilled, Tap, Waste and Seawater

1989 ◽  
Vol 21 (3) ◽  
pp. 255-258 ◽  
Author(s):  
Evangélos Biziagos ◽  
Jacques Passagot ◽  
Jean-Marc Crance ◽  
Robert Deloince
Keyword(s):  
Cell Culture ◽  
Polyethylene Glycol ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Tap Water ◽  
Virus Concentration ◽  
Beef Extract ◽  
Polyethylene Glycol 6000

The concentration of cell-culture-adapted hepatitis A virus (HAV) from experimentally contaminated distilled, drinking, waste and seawater was performed by using a filter adsorption-elu-tion method in the following conditions: HAV seeded in water was adsorbed at pH 4.0 to two nitrocellulose membranes (1.2 and 0.45 µm porosity for distilled and tap water or 8.0 and 3.0 µm porosity for waste and seawater), then eluted by 3% beef-extract at pH 8.5 and further concentrated by polyethylene glycol 6000 precipitation. Thus, HAV in 5 to 50 liters of seeded waters was concentrated approximately 1,700 to 17,000 fold with greater than 70% recovery of the initial virus added to the samples.

Epidemiologic Patterns of Wild-Type Hepatitis A Virus Determined by Genetic Variation

1991 ◽  
Vol 163 (2) ◽  
pp. 286-292 ◽  
Author(s):  
B. H. Robertson ◽  
B. Khanna ◽  
O. V. Nainan ◽  
H. S. Margolis
Keyword(s):  
Genetic Variation ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Wild Type

In Vivo Replication and Reversion to Wild Type of a Neutralization-Resistant Antigenic Variant of Hepatitis A Virus

1990 ◽  
Vol 161 (1) ◽  
pp. 7-13 ◽  
Author(s):  
S. M. Lemon ◽  
L. N. Binn ◽  
R. Marchwicki ◽  
P. C. Murphy ◽  
L.-H. Ping ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Wild Type ◽  
Antigenic Variant

scholarly journals Quantification of Hepatitis A Virus in Shellfish by Competitive Reverse Transcription-PCR with Coextraction of Standard RNA

1999 ◽  
Vol 65 (1) ◽  
pp. 322-326 ◽  
Author(s):  
Charlotte Arnal ◽  
Virginie Ferre-Aubineau ◽  
Berangere Mignotte ◽  
Berthe Marie Imbert-Marcille ◽  
Sylviane Billaudel
Keyword(s):  
Tissue Culture ◽  
Reverse Transcription ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Internal Standard ◽  
Wild Type ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Reproducible Method ◽  
Dna Enzyme Immunoassay

ABSTRACT To quantify hepatitis A virus (HAV) in experimentally contaminated mussels, we developed an internal standard RNA with a 7-nucleotide deletion for competitive reverse transcription (RT)-PCR. Deposited directly into the sample, this standard was used both as extraction control and as quantification tool. After coextraction and competitive RT-PCR, standard and wild-type products were detected by differential hybridization with specific probes and a DNA enzyme immunoassay. The quantifiable range with this reproducible method was 104 to 107 copies of HAV/gram or 400 to 106 50% tissue culture infective doses/ml.

Survival of Hepatitis A Virus in Spinach during Low Temperature Storage

2009 ◽  
Vol 72 (11) ◽  
pp. 2390-2393 ◽  
Author(s):  
Y. CAROL SHIEH ◽  
DIANA S. STEWART ◽  
DAVID T. LAIRD
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Plaque Assay ◽  
Storage Conditions ◽  
Inactivation Rate ◽  
Low Temperature Storage ◽  
Foodborne Outbreaks ◽  
Phosphate Buffered Saline ◽  
Temperature Storage ◽  
Spinach Leaves

Spinach leaves are frequently consumed raw and have been involved with past foodborne outbreaks. In this study, we examined the survival of hepatitis A virus (HAV) on fresh spinach leaves in moisture- and gas-permeable packages that were stored at 5.4 ± 1.2°C for up to 42 days. Different eluents including phosphate-buffered saline (PBS), pH 7.5 (with and without 2% serum), and 3% beef extract (pH 7.5 and 8) were compared for how efficiently they recovered viruses from spinach by using a simple elution procedure (<1 h). The recoveries were compared and determined by a plaque assay with FRhK-4 cells. Culture grade PBS containing 2% serum was found to be appropriate for HAV elution from spinach leaves, with an average recovery of 45% ± 10%. Over 4 weeks of storage at 5.4 ± 1.2°C, HAV in spinach decreased slightly more than 1 log, with 6.75% of the original titer remaining. HAV survived under refrigerated temperatures on spinach leaves with a D-value of 28.6 days (equivalent to an inactivation rate of 20.035 log of HAV per day, r2 = 0.88). In comparison, HAV in PBS containing 2% serum under the same storage conditions remained constant throughout 7 weeks. The inactivation rate of 20.035 log each day for HAV on spinach leaves was possibly due to the interaction of the virus and the leaf.

scholarly journals Development and Evaluation of a Broadly Reactive TaqMan Assay for Rapid Detection of Hepatitis A Virus

2005 ◽  
Vol 71 (6) ◽  
pp. 3359-3363 ◽  
Author(s):  
N. Jothikumar ◽  
T. L. Cromeans ◽  
M. D. Sobsey ◽  
B. H. Robertson
Keyword(s):  
Rapid Detection ◽  
Environmental Samples ◽  
Untranslated Region ◽  
Hepatitis A ◽  
Screening Tool ◽  
Hepatitis A Virus ◽  
Taqman Probe ◽  
Taqman Assay

ABSTRACT Primers and a TaqMan probe for the 5′-untranslated region (UTR) of the hepatitis A virus (HAV) genome were designed and evaluated. The assay detected 0.5 infectious units of HAV and 40 copies of a synthetic transcript and provides an important screening tool for rapid quantitative HAV detection in clinical or environmental samples.

Development of a plaque assay for a cytopathic, rapidly replicating isolate of hepatitis A virus

1987 ◽  
Vol 22 (1) ◽  
pp. 45-56 ◽  
Author(s):  
Theresa Cromeans ◽  
Mark D. Sobsey ◽  
Howard A. Fields
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Plaque Assay
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