Polyacrylamide gel electrophoresis of RNA compared with polyclonal- and monoclonal-antibody-based enzyme immunoassays for rotavirus
| Autor: | Vincent V. Hamparian, M J Connell, C T Budde, John H. Hughes, D L Pacini, Michael T. Brady |
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| Rok vydání: | 1988 |
| Předmět: |
Diarrhea
Rotavirus Microbiology (medical) medicine.drug_class Antibodies Viral Monoclonal antibody medicine.disease_cause Rotavirus Infections Immunoenzyme Techniques fluids and secretions Predictive Value of Tests medicine Humans Prospective Studies Child Polyacrylamide gel electrophoresis Cross Infection biology Antibodies Monoclonal RNA Virology Molecular biology Polyclonal antibodies Monoclonal biology.protein RNA Viral Electrophoresis Polyacrylamide Gel Reagent Kits Diagnostic Antibody medicine.symptom Research Article |
| Zdroj: | Journal of Clinical Microbiology. 26:194-197 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.26.2.194-197.1988 |
| Popis: | Polyacrylamide gel electrophoresis (PAGE) of rotaviral RNA, a sensitive and highly specific test for detecting rotavirus in stool, was compared with two commercially available enzyme immunoassays (EIAs), monoclonal (Pathfinder) and polyclonal (Rotazyme II). Stool samples from 204 children with nosocomial diarrhea were tested for rotavirus by both EIAs and by PAGE of RNA extracted from raw stools or 10% stool suspensions. Samples which tested positive by either EIA but were negative by PAGE were subjected to blocking EIA with rabbit or goat anti-SA11. Rotavirus was detected by PAGE and Pathfinder in 62 stools, but only 47 of these were positive by Rotazyme II. Blocking assays of EIA-positive, PAGE-negative samples suggested the presence of rotavirus in four additional stools. Sensitivity and specificity measured against PAGE and blocking assays were: Pathfinder, 0.985 and 0.934; and Rotazyme II, 0.731 and 0.927, respectively. False-positive rates were 0.134 for Pathfinder and 0.149 for Rotazyme II. The specificity and rate of false-positive results of Pathfinder were improved by using an adjusted optical density cutoff 4.36 times greater than that recommended by the manufacturer (specificity, 0.993; sensitivity, 0.985; false-positive rate, 0.015). |
| Databáze: | OpenAIRE |
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