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.

Detection of Hepatitis a Virus in Drinking Water by Enzyme -Immunoassay Using Ultracentrifugation for Virus Concentration

1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger
Keyword(s):  
Membrane Filtration ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Phosphate Buffer Solution ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Concentration ◽  
Buffer Solution ◽  
Solution Ph ◽  
Two Stage

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.

Restricted replication of human hepatitis A virus in cell culture: intracellular biochemical studies.

1981 ◽  
Vol 37 (1) ◽  
pp. 216-225 ◽  
Author(s):  
S A Locarnini ◽  
A G Coulepis ◽  
E G Westaway ◽  
I D Gust
Keyword(s):  
Cell Culture ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Human Hepatitis ◽  
Biochemical Studies

Hepatitis A virus cDNA and its RNA transcripts are infectious in cell culture.

1987 ◽  
Vol 61 (10) ◽  
pp. 3035-3039 ◽  
Author(s):  
J I Cohen ◽  
J R Ticehurst ◽  
S M Feinstone ◽  
B Rosenblum ◽  
R H Purcell
Keyword(s):  
Cell Culture ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Rna Transcripts ◽  
Virus Cdna

Mutational Events in Consecutive Passages of Hepatitis A Virus Strain GBM during Cell Culture Adaptation

Virology ◽  
1994 ◽  
Vol 204 (1) ◽  
pp. 60-68 ◽  
Author(s):  
Judith Graff ◽  
Christa Kasang ◽  
Andrea Normann ◽  
Mechtild Pfisterer-Hunt ◽  
Stephen M. Feinstone ◽  
...  
Keyword(s):  
Cell Culture ◽  
Virus Strain ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Culture Adaptation

Viral Enhancement and Interference Induced in Cell Culture by Hepatitis A Virus: Application to Quantitative Assays for Hepatitis A Virus

1984 ◽  
Vol 175 (1) ◽  
pp. 84-87
Author(s):  
W. M. Hurni ◽  
W. J. Miller ◽  
W. J. McAleer ◽  
P. J. Provost ◽  
M. R. Hilleman
Keyword(s):  
Cell Culture ◽  
Hepatitis A ◽  
Hepatitis A Virus
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