Comparison of five diagnostic methods for detecting bovine viral diarrhea virus infection in calves.
Five diagnostic techniques performed on skin biopsies (shoulder region) and/or serum were compared for detection of bovine viral diarrhea virus infection in 224 calves 0-3 months of age, 23 calves older than 3 months but younger than 7 months, and 11 cattle older than 7 months. The diagnostic methods used were immunohistochemistry (IHC), 2 commercial antigen ELISAs, 1 commercial antibody ELISA, and real-time RT-PCR. Results of 249 out of 258 skin and serum samples were identical and correlated within the 3 antigen detection methods and the real-time RT-PCR used. Twenty-six of these 249 samples were BVDV-positive with all antigen detection methods and the real-time RT-PCR. Nine out of 258 samples yielding discordant results were additionally examined by RT-PCR, RT-PCR Reamplification (ReA), and antigen ELISA I on serum and by immunohistochemistry on formalin fixed and paraffin-embedded skin biopsies. Virus isolation and genotyping was performed as well on these discordant samples. In 3 cases, transiently infected animals were identified. Two samples positive by real-time RT-PCR were interpreted as false positive and were ascribed to cross-contamination. The antigen ELISA II failed to detect 2 BVDV-positive calves due to the presence of maternal antibodies; the cause of 2 false-positive cases in this ELISA remained undetermined. Only persistently infected animals were identified in skin samples by IHC or antigen ELISA I. The 3 antigen detection methods and the real-time RT-PCR used in parallel had a high correlation rate (96.5%) and similar sensitivity and specificity values.
Hilbe
Monika
M
Institute of Veterinary Pathology, Winterthurerstrasse 268, 8057 Zurich, University of Zurich, Vetsuisse-Faculty, Switzerland. hilbe@vetpath.unizh.ch
Stalder
Hanspeter
H
Peterhans
Ernst
E
Haessig
Michael
M
Nussbaumer
Marlies
M
Egli
Christoph
C
Schelp
Christian
C
Zlinszky
Kati
K
Ehrensperger
Felix
F
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000175:diagnosis* / Q000821:virology*; D002417:Cattle; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012867:Skin / Q000276:immunology
D000914:Antibodies, Viral