Viral intestinal infections of animals and man

Eugster, Anne; Sneed, Loyd

doi:10.1016/0147-9571(79)90084-5

Cited by 11 publications

(10 citation statements)

References 36 publications

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“…27 Detection of the pleomorphic coronavirus particles in clinical specimens, such as respiratory secretions and stools, was initially done by DEM, although failure to preserve surface projections and the presence of particles that morphologically resemble coronaviruses make DEM less suitable for precise identification of coronavirus-positive or -negative specimens. 10,11 Prior investigations confirmed that when fecal samples containing BCV or TCV were examined by DEM coronavirus particles were usually widely spaced and the background heavily contaminated with debris. 3,5 Fringed particles that could be mistaken for coronaviruses were frequently observed, particularly in cases of specimens obtained from diarrheic turkey poults.…”

Section: Discussionmentioning

confidence: 99%

“…Coronaviruses are difficult to identify in negatively stained preparations because they are very pleomorphic and the peplomers are lost during processing of clinical specimens. 10,11,14 It is difficult to distinguish coronavirus particles from cell artifacts such as parts of nuclear or cytoplasmic membranes and other fringed bodies 10 Several immunoelectron microscopy (IEM) . methods for detecting enteric coronaviruses are more sensitive than conventionnal DEM.…”

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confidence: 99%

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Identification of Coronaviruses by the Use of Indirect Protein A-Gold Immunoelectron Microscopy

Dea

Garzón

1991

J VET Diagn Invest

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Abstract. Concentration by airfuge and protein A-colloidal gold immunoelectron microscopy (PAG-IEM) offered a rapid and sensitive method for detection and identification of coronaviruses from various species. The method was applied to partially purified tissue culture-adapted or egg-adapted mammalian and avian coronaviruses and to clarified fecal samples from diarrheic calves and turkey poults for detection of enteric coronaviruses. Aggregates of virus coated with specific antibody were seen in virus samples mixed with homologous antiserum but not in control samples containing preexposure serum. At least a 10-50-fold enhancement of the sensitivity of direct EM for virus detection was obtained using protein A-colloidal gold complex as an electron-dense marker. The PAG-IEM method demonstrated low nonspecific background labeling and permitted detection of soluble and particle-associated antigen. Reciprocal cross-reactivity was detected among the subgroup of mammalian hemagglutinating coronaviruses, and antisera to 4 members of other subgroups only recognized their homologous virus.Coronaviruses, particularly enteropathogenic strains, from avian and mammalian species are difficult to grow in cell cultures. 2 6 Direct electron microscopy (DEM) of viral particles in clarified fecal specimens and fluorescent antibody techniques for detection of coronavirus antigens in the cells of infected gut are generally used for the diagnosis of enteric coronaviral infections. 11,14,18,19,21 The detection of coronavirus antigens by indirect immunofluorescence (IFF) is dependent on tissue being removed very shortly after death, and most antigen is present early in the course of disease. 19,21 The lack of highly specific antisera can also compromise the specificity of the IFF technique. Coronaviruses are difficult to identify in negatively stained preparations because they are very pleomorphic and the peplomers are lost during processing of clinical specimens. 10,11,14 It is difficult to distinguish coronavirus particles from cell artifacts such as parts of nuclear or cytoplasmic membranes and other fringed bodies 10 Several immunoelectron microscopy (IEM) . methods for detecting enteric coronaviruses are more sensitive than conventionnal DEM. techniques; 17,24 however, the sensitivity of these methods is still dependent on the morphologic identification of virus particles or immune complexes on the EM grids. The recently described colloidal gold immunoelectron mi- Presented at the 33rd Annual Meeting of the AAVLD, Denver, CO, October 6-11, 1990.Received for publication February 13, 1991. croscopy technique improves detection of bovine enteric coronavirus (BCV) in fecal samples. 9 The purpose of the present study was to evaluate the efficiency of combining the concentration of viral suspensions by airfuge and the indirect protein A-colloidal gold immunoelectron microscopy labeling (PAG-IEM) techniques for the detection and differentiation of avian and mammalian enteric and respiratory coronaviruses. Materials and methodsViruses an...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Identification of Coronaviruses by the Use of Indirect Protein A-Gold Immunoelectron Microscopy

Dea

Garzón

1991

J VET Diagn Invest

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“…Second, small CVLPs were observed in the process of budding-off from larger fringed particles by a narrow stalk. Coronavirus-like particles also tend to exclude the negative stain, whereas nonspecific membrane profiles are penetrated by stain (Eugster and Sneed, 1980). Third, CVLPs were noted to be morphologically different from nonspecific membranes.…”

Section: Complement-like Lesionsmentioning

confidence: 98%

An Eight-Year Study of the Viral Agents of Acute Gastroenteritis in Humans: Ultrastructural Observations and Seasonal Distribution with A Major Emphasis on Coronavirus-like Particles

Borduin

et al. 1987

Coronaviruses

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During an 8-yr period, 862 stool specimens from patients with gastroenteritis were examined by electron microscopy after negative staining with 2% phosphotungstic acid (pH 6.5). Fortyone percent of the specimens submitted over an 8-yr period were determined to be positive for virus or viruslike particles belonging to one or more of seven morphologically distinct viral groups. Coronavirus-like particles (CVLPs) were present in 69.8% of the positive stool specimens. Membranous profiles containing "complement-type" holes (10 nm in diameter) were identified in some preparations containing CVLPs. The second most prevalent viral agent found in stool specimens was the rotavirus (17% of all positive stools). The incidence of other viruses identified in the survey were as follows: adenovirus 4.5%, picorna/parvovirus agents 2.9%, Norwalk-like agent 2.9%, astrovirus 1.9%, and calicivirus 0.5%. Unclassified small round viruses (-25-30 nm in diameter) represented 0.5%. It was also determined that there was a seasonal distribution in excretion of all viruses except for CVLPs. A greater number of viruses were identified in the cooler, drier months of the year.

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“…Thin sectioning and negatively stained transmission electron microscopy was performed as described [4,6]. The effects of lipid solvents, nucleic acid determination, heat and acid stability, and the effect of divalent cations were studied [2].…”

Section: Virus Characterizationmentioning

confidence: 99%

Isolation and characterization of caliciviruses from dogs with vesicular genital disease

Crandell

1988

Archives of Virology

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Two virus isolates, one from lesions of the vagina of a Bearded Collie and the other from the penis and prepuce of a Black Labrador, were partially characterized. The two viruses possessed the physicochemical properties, size and morphology of viruses belonging to the family Caliciviridae. The two isolates were shown by cross neutralization tests to be distinct from previously reported canine and feline caliciviruses. The viruses, isolated four years apart, are antigenically related. Additional studies are necessary to determine whether they are two distinct viruses or strains of another serotype belonging to the caliciviruses of the canine species.

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Viral intestinal infections of animals and man

Cited by 11 publications

References 36 publications

Identification of Coronaviruses by the Use of Indirect Protein A-Gold Immunoelectron Microscopy

Identification of Coronaviruses by the Use of Indirect Protein A-Gold Immunoelectron Microscopy

An Eight-Year Study of the Viral Agents of Acute Gastroenteritis in Humans: Ultrastructural Observations and Seasonal Distribution with A Major Emphasis on Coronavirus-like Particles

Isolation and characterization of caliciviruses from dogs with vesicular genital disease

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