title: Diagnostic performance of ELISA and PCR in identifying SRLV-infected sheep and goats using serum, plasma and milk samples and in early detection of infection in dairy flocks through bulk milk testing.
abstract: The objective of the study was to evaluate the diagnostic performances of the ELITEST-MVV ELISA for detection of antibodies against small ruminant lentiviruses and of two recently published PCRs for the detection of proviral DNA of SRLV in blood and corresponding individual milk samples. In addition, the feasibility of bulk milk testing was investigated by titrating ELISA positive pooled milk samples in negative milk, and by investigating bulk milk samples by ELISA and PCR in relation to the SRLV-status of the flocks. The results show that plasma and milk are suitable replacements for serum. For sheep, both PCRs showed a better diagnostic performance than for goats. ELISA results for bulk milk samples were promising with a putative detection limit of <3% within-herd prevalence using 1/10 pre-diluted samples and even <1% within-herd prevalence when samples were tested undiluted. In a panel of 249 bulk milk samples, all samples from SRLV free flocks (n=138) tested negative in the ELISA, while 50% of the samples from flocks with an unknown SRLV-status (n=111) were positive. For a subset of 59 bulk milk samples, agreement between ELISA results and leader-gag PCR results was almost 100%. These results demonstrate the potential of bulk milk testing as a cost effective tool for early detection of infection in dairy flocks, which is essential for SRLV-monitoring programs.
authors.affiliation: Department for Diagnostics, Research and Epidemiology, Animal Health Service Deventer, Deventer, The Netherlands. j.brinkhof@gddeventer.com
authors.forename: J M A
authors.initials: JM
authors.lastname: Brinkhof
authors.forename: D J
authors.initials: DJ
authors.lastname: Houwers
authors.forename: L
authors.initials: L
authors.lastname: Moll
authors.forename: D
authors.initials: D
authors.lastname: Dercksen
authors.forename: C
authors.initials: C
authors.lastname: van Maanen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood; D016088:Arthritis-Encephalitis Virus, Caprine / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D015511:Goat Diseases / Q000175:diagnosis*; D006041:Goats; D016180:Lentivirus Infections / Q000175:diagnosis / Q000662:veterinary*; D057230:Limit of Detection; D008892:Milk / Q000737:chemistry / Q000276:immunology; D011021:Pneumonia, Progressive Interstitial, of Sheep / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011159:Population Surveillance / Q000379:methods; D012367:RNA, Viral / Q000032:analysis; D015203:Reproducibility of Results; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis*; D014790:Visna-maedi virus / Q000502:physiology
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral