Agarose Gel Filtration of Concentrated Fecal Extracts Containing Virus-Like Particles Associated with Hepatitis A

Cl, Hornbeck; Cr, Gravelle; Fb, Hollinger; Dw, Bradley

doi:10.1159/000149484

Cited by 9 publications

(3 citation statements)

References 13 publications

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“…Hornbeck et al [23] purified HAV from the feces of infected chimpanzees by a combination of polyethylene glycol (PEG) precipitation, isopycnic banding in CsCl gradients, and aga rose gel filtration using Scpharose 2B. These procedures removed HAV from more than 90% of extraneous fecal material [24], and the addition of 0.5% (w/v) bovine serum albumin…”

Section: Purification O F Ha V From Fecal Specimensmentioning

confidence: 99%

Biophysical and Biochemical Characterization of Hepatitis A Virus

et al. 1982

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Biophysical and biochemical analysis of hepatitis A virus has shown it to be a 27- to 32-nm icosahedral particle with 32 capsomers. The mature virion has a buoyant density of 1.33–1.34 g/cm³, a sedimentation coefficient of 156–160S, and is composed of four polypeptides with molecular weights of 30,000–33,000 (VPl), 24,000–27,000 (VP2), 21,000–23,000 (VP3), and 7,000–14,000 (VP4). The genome of hepatitis A virus consists of a single piece of single-stranded RNA which sediments at 32–35S and has a buoyant density of 1.64 g/cm³. The molecular weight of RNA is 2.25 × 10⁶ when measured under nondenaturing conditions and 2.8 × 10⁶ when measured under fully denaturing conditions. The genome contains a 40–80 nucleotide sequence of polyadenylic acid and is capable of infecting cell cultures. These findings, together with the observation that the virion is stable at pH 3.0 and resistant to ether and a temperature of 60° for 1 h, indicate that hepatitis A virus should now be classified as an Entero-virus within the family Picornaviridae.

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Section: Purification O F Ha V From Fecal Specimensmentioning

confidence: 99%

Biophysical and Biochemical Characterization of Hepatitis A Virus

et al. 1982

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“…Chloroform extraction appears to be more effective than fluorocarbon, but has been shown to damage some entero viruses [9], The use of Sepharose CL-2B to remove extraneous fecal material from HAV was first reported by Hornbeck el at. [10]. In a previous study, we found that this procedure was capable of removing 90 99% of extraneous material absorbing at 280 nm [7].…”

Section: Discussionmentioning

confidence: 91%

Purification of Hepatitis A Virus from Human Feces

et al. 1978

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Hepatitis A virus was purified from fecal specimens obtained from 2 patients with naturally acquired hepatitis A. The purification procedure involved differential centrifugation, organic solvent extraction, agarose gel filtration, ion-exchange chromatography, and isopycnic ultracentrifugation in cesium chloride. Using immune electron microscopy and discontinuous SDS-PAGE, this procedure was found to be effective in removing extraneous material from hepatitis A virus. There was significant recovery of virus as judged by immune electron microscopy and solid-phase radioimmunoassay. Using this protocol, it has been possible to obtain virus preparations of sufficient purity and high enough titer to enable biochemical studies to proceed.

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“… 2 The sedimentation coefficiency of this virus is calculated to be 57–60 S. It has a density of 1.33 g/cm 3 in a CsCl solution and 1.34 g/cm 3 in a neutral sucrose solution. 9 Proteins with molecular weights of 30 000–33 000, 24 000–27 000, 21 000–23 000 and 7000–14 000 Da were identified in viral particles 17–20 and, when compared with other picorna viruses, these were considered to be capsid proteins: VPl, VP2, VP3 and VP4, respectively. However, a subsequent study disputed the presence of VP4.…”

Section: Physicochemical Structure Of Type a Hepatitis Virusmentioning

confidence: 99%

Molecular biology of hepatitis A virus: Significance of various substitutions in the hepatitis A virus genome

Yokosuka

2000

J of Gastro and Hepatol

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Hepatitis A virus (HAV) is the sole member of the hepatogenus of Picorna viridae. This virus can now be propagated in cell culture and in primates. Molecular biological studies of HAV have disclosed its genomic structure and the functional significance of the viral proteins to some extent. Hepatitis A virus has a positive‐stranded RNA of approximately 7.5 kb that encodes a large polyprotein. Translation of the protein is influenced by the function of the internal ribosomal entry site in the 5′ non‐translating region. It is generally agreed that the polyprotein is processed to four structural and seven non‐structural proteins by the proteinase encoded in the 3C region. Replication efficiency seems to be controlled by amino acid substitutions in the 2B and 2C regions. The virulence of HAV in primates may be determined by substitutions in the 2C region. Although the severity of hepatitis A was thought to be determined by immunological reactions of the host to the virus, the potential virulence of the variant viruses themselves may need further examination. Recent progress in polymerase chain reaction technology has made possible an analysis of the HAV sequence in clinical specimens; such analysis is of importance in the disclosure of differences in HAV subspecies in different clinical conditions.

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Agarose Gel Filtration of Concentrated Fecal Extracts Containing Virus-Like Particles Associated with Hepatitis A

Cited by 9 publications

References 13 publications

Biophysical and Biochemical Characterization of Hepatitis A Virus

Biophysical and Biochemical Characterization of Hepatitis A Virus

Purification of Hepatitis A Virus from Human Feces

Molecular biology of hepatitis A virus: Significance of various substitutions in the hepatitis A virus genome

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