Development of a 1-step enzyme-linked immunosorbent assay for the rapid diagnosis of bovine respiratory syncytial virus in postmortem specimens.
Bovine respiratory syncytial virus (BRSV) is associated with severe respiratory disease in cattle. BRSV infection frequently leads to the death of young infected animals. The presence of BRSV in postmortem specimens is routinely detected using indirect immunofluorescence (IIF). However, this technique requires special equipment and considerable expertise. The present paper describes the development of a 1-step ELISA for rapid (1.5 hours) detection of BRSV antigen in organ homogenates. The performance of the new 1-step ELISA was evaluated using bovine postmortem specimens (n = 108) in comparison with 3 other BRSV diagnostic techniques: indirect immunofluorescence, the Clearview respiratory syncytial virus (RSV) test, and real-time reverse transcriptase polymerase chain reaction (RT-PCR). The relative sensitivity, specificity, and the kappa coefficient of 1-step ELISA, the Clearview RSV electroimmunoassay (EIA), and IIF were calculated, using real-time RT-PCR as the reference test. The new 1-step ELISA was the most sensitive and specific of the 3 tests. Thus, the new 1-step ELISA is a reliable test for detecting BRSV antigen in organ homogenates.
Quinting
Birgit
B
Centre d'Economie Rurale, Division Immunologie Animale, rue du Carmel, 1, 6900 Marloie, Belgium. b.quinting@cergroupe.be
Robert
Béatrice
B
Letellier
Carine
C
Boxus
Mathieu
M
Kerkhofs
Pierre
P
Schynts
Frédéric
F
Collard
Alfred
A
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D007124:Immunoenzyme Techniques / Q000662:veterinary; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D015203:Reproducibility of Results; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D018112:Respiratory Syncytial Virus, Bovine / Q000235:genetics / Q000302:isolation & purification*; D012140:Respiratory Tract Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity
D000956:Antigens, Viral; D012367:RNA, Viral