Improvement of rotavirus isolation in the cell culture by immune peroxidase staining

Weber, Bernard; Harms, F.; Selb, B.; Doerr, H. W.

doi:10.1016/0166-0934(92)90109-q

Cited by 6 publications

(3 citation statements)

References 21 publications

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“…Although the efficiency of rotavirus isolation has been improved by spin amplification technique and detection of rotavirus antigen by monoclonal antibody coupled to immunoperoxidase staining [10] or by fluorescent focus assay [15], the sensitivity is still under that of current commercial EIAs [10], and the method is relatively labor intensive, so that it is not suited for the evaluation of a large number of specimens.…”

Section: Discussionmentioning

confidence: 99%

“…The inoculated wells were incubated for 1 h at 37°, and the supernatant was replaced by maintenance medium. The immune peroxidase staining was performed according to the method described by Weber et al [10] for the detection of rotavirus. Briefly, maintenance medium was aspirated and the cells were fixed with 100 Ìl of ice-cold acetone/methanol (2:3) during 10 min.…”

Section: Immunoperoxidase Staining For the Detection Of Adenovirus Inmentioning

confidence: 99%

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Improvement of the Specificity of Enzyme Immunoassays for the Detection of Rotavirus and Adenovirus in Fecal Specimens

Rabenau¹,

Knoll²,

Allwinn³

et al. 1998

Intervirology

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A variable rate of false-positive results may be observed with commercial enzyme immunoassays (EIAs) for the detection of rotavirus and adenovirus antigen in stool specimens, depending on the quality of the reagents and the presence of potentially interfering substances in stool samples. The present study was performed in an attempt to improve the specificity of current commercial rotavirus and adenovirus EIAs without significant loss of sensitivity by optimizing the cut-off value. A collective of 174 stool samples obtained from children suffering from acute gastroenteritis was tested. Electron microscopy (EM) and PAGE were used as reference methods for rotavirus detection. For the evaluation of the adenovirus kits, virus isolation in cell culture and the polymerase chain reaction served as reference standards. The highest sensitivity for rotavirus and adenovirus detection was achieved by the Ridascreen^® Rotavirus and Ridascreen^® Adenovirus. However, the Ridascreen^® Rotavirus and Ridascreen^® Adenovirus produced the highest number of false-positive results (n = 9) for each rotavirus and adenovirus detection. Cross-reactivities to coronaviruses and reoviruses were observed with the rotavirus antigen EIAs. For Rotazyme II, Ridascreen Rotavirus and Ridascreen Adenovirus, the specificity could be markedly increased without loss of sensitivity by doubling the cut-off value. For the alternative immunoassays, which were overall more specific, it was not possible to significantly decrease the rate of false-positive results without impairment of sensitivity by raising the cut-off value. In conclusion, at least for some rotavirus and adenovirus antigen EIAs, the cut-off value set by the manufacturer may not permit an optimal differentiation between true-positive and -negative samples. By raising the cut-off value from 50 to 100%, the specificity of two rotavirus antigen and one adenovirus antigen EIA can be improved markedly without significant loss of sensitivity.

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Section: Discussionmentioning

confidence: 99%

Section: Immunoperoxidase Staining For the Detection Of Adenovirus Inmentioning

confidence: 99%

Improvement of the Specificity of Enzyme Immunoassays for the Detection of Rotavirus and Adenovirus in Fecal Specimens

Rabenau¹,

Knoll²,

Allwinn³

et al. 1998

Intervirology

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“…In contrast, rotavirus was isolated from only 4 (3.3%) fecal specimens by conventional cell culture (i.e. demonstration of a cytopathogenic effect) while a total of 93 (76.9%) samples were positive by ELISA [65].…”

Section: Genes Reviewmentioning

confidence: 93%

Evaluation of Techniques for the Diagnosis of Rotavirus Gastroenteritis in Paediatric Patients

R.K¹,

Oke²,

Anjorin³

et al. 2015

Genes Review

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Rotavirus is the major etiology of acute gastroenteritis in infants and young children worldwide. It has been estimated that about 39% of childhood diarrhoea hospitalizations are caused by the virus and nearly half a million children die from the infections each year especially in developing countries on the Asian subcontinent, Africa, and Latin America where health care facilities are in short supply. Stool specimens collected from the first to fourth days of illness are optimal for detection of the infection. Diagnosis was originally performed using electron microscopy, which is still occasionally used in centers where it is available. Routine diagnosis is however, now performed by antigen detection on feces using commercially available, simple, rapid immunochromatographic dipstick style kits which have superseded the earlier latex agglutination and enzyme immunoassay methods whose usefulness in diagnosis is still relevant. Reverse transcription polymerase chain reaction of feces is also available in some reference and research centers, and is particularly useful for identification of outbreaks due to serogroups other than group A. Similarly viral culture and serology are also available but do not play much roles in diagnosis of acute disease. Rotavirus infection is indeed a public health problem and early, rapid, accurate and reliable diagnostic techniques are essential for effective patient management and infection control.

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Processing of Tissue Culture Cells

Bratthauer¹

2009

Immunocytochemical Methods and Protocols

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Improvement of rotavirus isolation in the cell culture by immune peroxidase staining

Cited by 6 publications

References 21 publications

Improvement of the Specificity of Enzyme Immunoassays for the Detection of Rotavirus and Adenovirus in Fecal Specimens

Improvement of the Specificity of Enzyme Immunoassays for the Detection of Rotavirus and Adenovirus in Fecal Specimens

Evaluation of Techniques for the Diagnosis of Rotavirus Gastroenteritis in Paediatric Patients

Processing of Tissue Culture Cells

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