Isolation of Single Stranded DNA from Purified Hepatitis A-Virus

Abstract Hepatitis A-Yirus-Single-Sranded D N A , Electron Microscopy, Length Measurements, Parvovirus Hepatitis A-virus was purified from human stools by three purification steps. Virus was identified by radioimmuno-assay and purity monitored with immune electron microscopy. Virus particles, serologically and morphologically identical, banded in CsCl in two density ranges at 1.31 — 1.34 g/cm3 and at 1.41 — 1.43 g/cm3. Virions of density 1.31 — 1.34 g/cm3 were shown to contain single-stranded D N A of different size classes. Class I 1.33 kb, class I I 4.61 kb in addition a small amount of molecules was de tected with lengths up to 15 kb.

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Application of solid-phase immune electron microscopy to study physical and stability characteristics of enterically transmitted non-a, non-b hepatitis virus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102481 ◽

1988 ◽

Vol 46 ◽

pp. 80-81

Author(s):

Charles D. Humphrey ◽

E. H. Cook ◽

Karen A. McCaustland ◽

Daniel W. Bradley

Keyword(s):

Electron Microscopy ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Solid Phase ◽

Etiologic Agent ◽

World Health ◽

Health Concern ◽

Morphological Identification ◽

Immune Electron Microscopy

Enterically transmitted non-A, non-B hepatitis (ET-NANBH) is a type of hepatitis which is increasingly becoming a significant world health concern. As with hepatitis A virus (HAV), spread is by the fecal-oral mode of transmission. Until recently, the etiologic agent had not been isolated and identified. We have succeeded in the isolation and preliminary characterization of this virus and demonstrating that this agent can cause hepatic disease and seroconversion in experimental primates. Our characterization of this virus was facilitated by immune (IEM) and solid phase immune electron microscopic (SPIEM) methodologies.Many immune electron microscopy methodologies have been used for morphological identification and characterization of viruses. We have previously reported a highly effective solid phase immune electron microscopy procedure which facilitated identification of hepatitis A virus (HAV) in crude cell culture extracts. More recently we have reported utilization of the method for identification of an etiologic agent responsible for (ET-NANBH).

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Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142815 ◽

1986 ◽

Vol 44 ◽

pp. 238-239

Author(s):

C.D. Humphrey ◽

T.L. Cromeans ◽

E.H. Cook ◽

D.W. Bradley

Keyword(s):

Electron Microscopy ◽

Liquid Phase ◽

Cell Cultures ◽

Rapid Detection ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Solid Phase ◽

Immune Electron Microscopy ◽

Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

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Hepatitis A Virus Aggregation in Suspensions of Purified Virus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010018015x ◽

1990 ◽

Vol 48 (1) ◽

pp. 280-281

Author(s):

Charles D. Humphrey ◽

Betty H. Robertson ◽

B. Khanna

Keyword(s):

Electron Microscopy ◽

Protein Function ◽

Hepatitis A ◽

Hepatitis A Virus ◽

X Ray ◽

Virus Particles ◽

X Ray Crystallography ◽

Morphological Studies ◽

Cryo Electron Microscopy ◽

Microscopic Evaluation

Virus capsid structure as determined by x-ray crystallography or by elec imaging of virus particles in thefrozen hydrated state has received con interest during the past decade. Specific functional proteins were localized recently on the Rotavirus capsid by using cryo-electron microscopy, indicating the potential for morphological studies that rel directly to capsid protein function.Most cryo-electron microscopy studies have involved investigation of vir were 40 nm or greater in diameter. Smaller viruses may provide a greate practical difficulty in their preparation and imaging.Highly purified virus for study by x-ray crystallographic studies or ele microscopic evaluation of frozen hydrated particles requires stringent c particle aggregation. Preparations for x-ray crystallography should agg an orderly manner to form arrays and ordered crystals of sufficient size Preparations for cryo-electron microscopy are more desirable when the pa numerous but not over-lapping (Fig. 1). We have experienced uncontrolla aggregation including considerable over-lapping of hepatitis A virus (HA particles when purified preparations were stored in phosphate or tris buffers (Fig. 2).

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Antibodies to hepatitis A virus: patterns by two procedures

Journal of Clinical Microbiology ◽

10.1128/jcm.5.1.110-111.1977 ◽

1977 ◽

Vol 5 (1) ◽

pp. 110-111

Author(s):

J Rakela ◽

D Stevenson ◽

V M Edwards ◽

I Gordon ◽

J W Mosley

Keyword(s):

Electron Microscopy ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Immune Electron Microscopy ◽

Immune Adherence ◽

Low Levels

Antibody to hepatitis A virus demonstrable by immune electron microscopy appeared early but remained at low levels for several weeks. Antibody detectable by immune adherence hemagglutination was delayed.

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Electron Microscope Study of RNA's of Paramyxoviruses

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094231 ◽

1972 ◽

Vol 30 ◽

pp. 196-197

Author(s):

Ruchama Baum ◽

J.T. Seto

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Electron Microscope Study ◽

Sendai Virus ◽

Sodium Dodecyl ◽

Rna Molecules ◽

Virus Particles ◽

Molecular Weights ◽

Length Measurements

The ribonucleic acid (RNA) of paramyxoviruses has been characterized by biochemical and physiochemical methods. However, paramyxovirus RNA molecules have not been studied by electron microscopy. The molecular weights of these single-stranded viral RNA molecules are not known as yet. Since electron microscopy has been found to be useful for the characterization of single-stranded RNA, this investigation was initiated to examine the morphology and length measurements of paramyxovirus RNA's.Sendai virus Z strain and Newcastle disease virus (NDV), Milano strain, were used. For these studies it was necessary to develop a method of extracting RNA molecules from purified virus particles. Highly purified Sendai virus was treated with pronase (300 μg/ml) at 37°C for 30 minutes and the RNA extracted by the sodium dodecyl sulfate (SDS)-phenol procedure.

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