Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Norwalk-like Viruses Associated With A Gastroenteritis Outbreak Following Oyster Consumption

1993 ◽  
Vol 56 (7) ◽  
pp. 604-607 ◽  
Author(s):  
RODERIC D. PONTEFRACT ◽  
F. R. BISHAI ◽  
J. HOCKEN ◽  
G. BERGERON ◽  
R. PARENT
Keyword(s):  
Electron Microscopy ◽  
Immunoelectron Microscopy ◽  
Solid Phase ◽  
Fecal Coliforms ◽  
Gastroenteritis Outbreak ◽  
Fecal Indicators ◽  
Negative Results ◽  
Thunder Bay ◽  
Stool Samples ◽  
Low Levels

In late October 1991, an outbreak of gastroenteritis, following the consumption of raw oysters involving more than 200 people, was reported in five locations in Quebec, Canada. Bacteriological analysis of the oysters involved indicated low levels of fecal coliforms, but direct electron microscopy of stool samples obtained from two people involved in the outbreak revealed that both contained 27–34 nm, small, round Norwalk-like viruses. Immunoelectron microscopy, using acute sera obtained from these individuals and convalescent serum from another person related to the outbreak, revealed antibody coatings on these Norwalk-like viruses with all three sera. Solid-phase immunoelectron microscopy demonstrated that these viruses were also antigenically similar or related to a Norwalk-like virus isolated as the cause of a gastroenteritis outbreak in a home for the aged between December 1988 and January 1989 in Thunder Bay, Ontario. From these findings and the symptoms of the illness, the Norwalk-like virus was considered as the causal agent of the outbreak due to the consumption of contaminated oysters. How the oysters became contaminated was not determined. Oysters harvested from the areas initially thought to have been the origin of the implicated shellfish were tested for the presence of viral fecal indicators using tissue culture and electron microscopy with negative results. It is most likely that the implicated lot also contained oysters harvested from another area, also open, but which was downstream from an identified source of human fecal contamination.

An investigation of experimental factors affecting the identification of enteroviruses by immunoelectron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 362-363
Author(s):  
F. K. Lee ◽  
N. Anderson ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Positive Result ◽  
Immunoelectron Microscopy ◽  
Cross Reactivity ◽  
Virus Antibody ◽  
Viral Isolate ◽  
Virus Diagnosis ◽  
Factors Affecting ◽  
Homologous Virus ◽  
Wide Range

In 1973 Anderson and Doane described a rapid method for serotyping enteroviruses on the EM grid, using immunoelectron microscopy (IEM). By mixing an unknown viral isolate with individual or pooled enterovirus antisera, the resulting typing test could be read by electron microscopy, using the presence of virus-antibody aggregates as indication of a positive result. The reaction was found to be serotype specific when tested with a wide range of enteroviruses (coxsackie, polio, echo). These results were in contrast to those obtained earlier by Chaudhary et al., who observed cross-reactivity among enteroviruses using IEM. In light of the potential usefulness of this method for rapid virus diagnosis, a study was carried out on some of the experimental factors that might affect the reliability of enterovirus serotyping by IEM.The most important factor in avoiding heterologous cross reactions was found to be the concentration of antiserum. As shown in Table I, when antisera were used at low dilutions they produced aggregation not only with the. homologous virus but also with the heterologous strains.

Ultrastructural investigation of spontaneous pituitary gonadotroph cell adenomas in aging Sprague-Dawley rats

1983 ◽  
Vol 41 ◽  
pp. 702-703
Author(s):  
D. J. McComb ◽  
J. Beri ◽  
F. Zak ◽  
K. Kovacs
Keyword(s):  
Electron Microscopy ◽  
Animal Model ◽  
Epoxy Resin ◽  
Immunoelectron Microscopy ◽  
Tumor Type ◽  
Gonadal Function ◽  
Sprague Dawley ◽  
Male And Female ◽  
Reticulin Fibers ◽  
Sprague Dawley Rats

Gonadotroph cell adenomas of the pituitary are infrequent in human patients and are not invariably associated with altered gonadal function. To date, no animal model of this tumor type exists. Herein, we describe spontaneous gonadotroph cell adenomas in old male and female Sprague-Dawley rats by histology, immunocytology and electron microscopy.The material consisted of the pituitaries of 27 male and 38 female Sprague Dawley rats, all 26 months of age or older, removed at routine autopsy. Sections of formal in-fixed, paraffin-embedded tissue were stained with hematoxylin-phloxine-saffron (HPS), the PAS method and the Gordon-Sweet technique for the demonstration of reticulin fibers. For immunostaining, sections were exposed to anti-rat β-LH, anti-ratβ-TSH, anti-rat PRL, anti-rat GH and anti-rat ACTH 1-39. For electron microscopy, tissue was fixed in 2.5% glutaraldehyde, postfixed in 1% OsO4 and embedded in epoxy-resin. Tissue fixed in 10% formalin, embedded in epoxy resin without osmification, was used for immunoelectron microscopy.

Ectodesmata as Revealed By Freeze-Etch Preparations of Plant Epidermal Cell Walls

1973 ◽  
Vol 31 ◽  
pp. 468-469
Author(s):  
N.C. Lyon ◽  
W. C. Mueller
Keyword(s):  
Electron Microscopy ◽  
Acetic Acid ◽  
Nitric Acid ◽  
Oxalic Acid ◽  
Light Microscopy ◽  
Epidermal Cell ◽  
Cell Walls ◽  
Plant Viruses ◽  
Plant Leaves ◽  
Thin Sections

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.

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.

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