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

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.

Download Full-text

Genetic variability of hepatitis A virus

Journal of General Virology ◽

10.1099/vir.0.19532-0 ◽

2003 ◽

Vol 84 (12) ◽

pp. 3191-3201 ◽

Cited By ~ 103

Author(s):

Mauro Costa-Mattioli ◽

Anna Di Napoli ◽

Virginie Ferré ◽

Sylviane Billaudel ◽

Raul Perez-Bercoff ◽

...

Keyword(s):

Genetic Variation ◽

Molecular Biology ◽

Genetic Variability ◽

Molecular Epidemiology ◽

Mutation Rate ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Genetic Recombination ◽

Classification Methods ◽

Evolutionary Mechanisms

Knowledge of the molecular biology of hepatitis A virus (HAV) has increased exponentially since its identification. HAV exploits all known mechanisms of genetic variation to ensure survival, including mutation and genetic recombination. HAV has been characterized by the emergence of different genotypes, three human antigenic variants and only one major serotype. This paper reviews the genetic variability and molecular epidemiology of HAV. Its evolutionary mechanisms are described with particular emphasis on genetic recombination and HAV mutation rate. Genotypic classification methods are also discussed.

Download Full-text

Complete nucleotide sequence of wild-type hepatitis A virus: comparison with different strains of hepatitis A virus and other picornaviruses.

Journal of Virology ◽

10.1128/jvi.61.1.50-59.1987 ◽

1987 ◽

Vol 61 (1) ◽

pp. 50-59 ◽

Cited By ~ 227

Author(s):

J I Cohen ◽

J R Ticehurst ◽

R H Purcell ◽

A Buckler-White ◽

B M Baroudy

Keyword(s):

Nucleotide Sequence ◽

Complete Nucleotide Sequence ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Wild Type ◽

Different Strains

Download Full-text

Relapsing hepatitis A in Saimiri monkeys experimentally reinfected with a wild type hepatitis A virus (HAV)

Chronically Evolving Viral Hepatitis - Archives of Virology ◽

10.1007/978-3-7091-5633-9_2 ◽

1992 ◽

pp. 5-10 ◽

Cited By ~ 5

Author(s):

S. Prevot ◽

J. Marechal ◽

J. Piilot ◽

J. Prevot

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Wild Type

Download Full-text

Concentration of Hepatitis a Virus in Environmental Samples

Water Science & Technology ◽

10.2166/wst.1991.0064 ◽

1991 ◽

Vol 24 (2) ◽

pp. 229-234 ◽

Cited By ~ 5

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.

Download Full-text

Complete nucleotide sequence of a cell culture-adapted variant of hepatitis a virus: Comparison with wild-type virus with restricted capacity for in vitro replication

Virology ◽

10.1016/0042-6822(88)90270-x ◽

1988 ◽

Vol 163 (2) ◽

pp. 299-307 ◽

Cited By ~ 64

Author(s):

Robert W. Jansen ◽

John E. Newbold ◽

Stanley M. Lemon

Keyword(s):

Cell Culture ◽

Nucleotide Sequence ◽

Complete Nucleotide Sequence ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Wild Type Virus ◽

Type Virus ◽

Wild Type ◽

A Cell

Download Full-text

Stable Growth of Wild-Type Hepatitis A Virus in Cell Culture

Journal of Virology ◽

10.1128/jvi.80.3.1352-1360.2006 ◽

2006 ◽

Vol 80 (3) ◽

pp. 1352-1360 ◽

Cited By ~ 43

Author(s):

Krishnamurthy Konduru ◽

Gerardo G. Kaplan

Keyword(s):

Cell Culture ◽

Cell Lines ◽

Hepatitis A ◽

Infectious Virus ◽

Hepatitis A Virus ◽

Wild Type ◽

Huh7 Cells ◽

Stable Growth ◽

I Cells

ABSTRACT Human wild-type (wt) hepatitis A virus (HAV), the causative agent of acute hepatitis, barely grows in cell culture and in the process accumulates attenuating and cell culture-adapting mutations. This genetic instability of wt HAV in cell culture is a major roadblock to studying HAV pathogenesis and producing live vaccines that are not overly attenuated for humans. To develop a robust cell culture system capable of supporting the efficient growth of wt HAV, we transfected different cell lines with in vitro RNA transcripts of wt HAV containing the blasticidin resistance gene. Blasticidin-resistant colonies grew only in transfected Huh7 cells and produced infectious virus. HAV was genetically stable in Huh7 cells for at least nine serial passages and did not accumulate attenuating or cell culture-adapting mutations. Treatment with alpha interferon A/D cured the blasticidin-resistant Huh7 cells of the HAV infection. The cured cells, termed Huh7-A-I cells, did not contain virus or HAV antigens and were sensitive to blasticidin. Huh7-A-I cells were more permissive than parental cells for wt HAV infection, including a natural isolate from a human stool sample, and produced 10-fold-more infectious particles. This is the first report of a cell line that allows the genetically stable growth of human wt HAV. The viral vectors and cells described here should allow better insight into the pathogenesis of HAV and the development of attenuated vaccines. The cell lines susceptible to wt HAV growth may also be used to detect and isolate infectious virus from patient and environmental samples.

Download Full-text