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

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).

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

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.

scholarly journals Enzyme-Linked Immunosorbent Assay for Detection of Hepatitis A Antigen in Stool and Antibody to Hepatitis A Antigen in Sera: Comparison with Solid-Phase Radioimmunoassay, Immune Electron Microscopy, and Immune Adherence Hemagglutination Assay

1978 ◽  
Vol 7 (2) ◽  
pp. 184-193
Author(s):  
Lars R. Mathiesen ◽  
Stephen M. Feinstone ◽  
Doris C. Wong ◽  
Peter Skinhoej ◽  
Robert H. Purcell
Keyword(s):  
Electron Microscopy ◽  
Hepatitis A ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immune Electron Microscopy ◽  
Specific Detection ◽  
Immune Adherence ◽  
Ag Particles ◽  
Solid Phase Radioimmunoassay ◽  
Human Stool

Previously described techniques for detection of hepatitis A antigen (HA Ag) and antibody (anti-HA) have required purified HA Ag and expensive equipment. Herein is described an enzyme-linked immunosorbent assay (ELISA) for specific detection of HA Ag in human stool filtrates and of anti-HA in sera by using selected HA Ag-containing human stool filtrates as the antigen source. Because human stools often react nonspecifically in serological tests for HA Ag, blocking with preexposure and hyperimmune anti-HA sera from a chimpanzee inoculated with hepatitis A virus was used to confirm specific detection of HA Ag. The sensitivity of ELISA was found to be comparable to that of solid-phase radioimmunoassay (SPRIA) and immune electron microscopy (IEM). Of 37 acute-phase stools collected from nine patients, 16 were positive for HA Ag by ELISA. In 13 of these, HA Ag particles were found by IEM, and an additional 3 stools negative by ELISA contained HA Ag particles by IEM. Eight control stools were negative by both ELISA and IEM. Anti-HA was measured in sera by demonstrating its ability to block binding of the enzyme conjugate to HA Ag in a stool without detectable nonspecificity. This test (blocking ELISA) was as sensitive and specific as blocking SPIRA, IEM, and immune adherence hemagglutination and, like SPRIA and IEM, detected early-developing antibody. The ELISA is simple to perform and requires only a minimum of equipment. It is useful for screening stools for HA Ag and for monitoring HA Ag during purification, as well as for detecting early and late anti-HA in sera.

scholarly journals Solid-phase enzyme-linked immunosorbent assay for detection of hepatitis A virus

1978 ◽  
Vol 8 (3) ◽  
pp. 277-282 ◽  
Author(s):  
S A Locarnini ◽  
S M Garland ◽  
N I Lehmann ◽  
R C Pringle ◽  
I D Gust
Keyword(s):  
Immunosorbent Assay ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Field Work ◽  
Enzyme Linked Immunosorbent Assay ◽  
Technical Equipment ◽  
Immune Electron Microscopy ◽  
Naked Eye ◽  
Household Contacts

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of hepatitis A virus in human fecal specimens. Investigations with 88 fecal specimens from 77 patients with suspected viral hepatitis and 8 of their household contacts showed that ELISA was as specific and sensitive as radioimmunoassay and almost as sensitive as immune electron microscopy. The ELISA is quick and simple to perform, does not require sophisticated technical equipment, and can be read with the naked eye, making it suitable for field work and rapid diagnosis.

Isolation of Single Stranded DNA from Purified Hepatitis A-Virus

1978 ◽  
Vol 33 (7-8) ◽  
pp. 594-597 ◽  
Author(s):  
Bertram Flehmig ◽  
Hans Dieter Royer ◽  
Hans-Joachim Gerth
Keyword(s):  
Electron Microscopy ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Class I ◽  
Immune Electron Microscopy ◽  
Single Stranded Dna ◽  
Virus Particles ◽  
Size Classes ◽  
Length Measurements

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.

scholarly journals Antibodies to hepatitis A virus: patterns by two procedures

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.

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.

The Relationship between a 27-nm Virus-Like Particle and Hepatitis A as Demonstrated by Immune Electron Microscopy

Intervirology ◽  
1974 ◽  
Vol 4 (2) ◽  
pp. 110-118 ◽  
Author(s):  
Stephen A. Locarnini ◽  
Allan A. Ferris ◽  
Anthony C. Stott ◽  
Ian D Gust
Keyword(s):  
Electron Microscopy ◽  
Hepatitis A ◽  
Immune Electron Microscopy ◽  
Virus Like Particle ◽  
The Relationship
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