Abstract. Isolation of Porcine reproductive and respiratory syndrome virus (PRRSV) from oral fluids was first reported in 1997. The objective of the present study was to determine whether PRRSV and/or anti-PRRSV antibodies were present in oral fluids at diagnostic levels. The level and duration of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in 3 age groups of pigs (4, 8, or 12 weeks of age) inoculated with a type 2 (North American) PRRSV isolate. Serum, buccal swabs, and pen-based oral fluid samples were collected for 63 days following inoculation. Specimens were assayed for PRRSV by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), and for anti-PRRSV antibodies by enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). Porcine reproductive and respiratory syndrome virus was detected by real-time qRT-PCR in serum for approximately 5 weeks and in oral fluids for approximately 4 weeks postinoculation. Pig age at the time of inoculation had no effect on the quantity or duration of virus in oral fluid samples. Low levels of anti-PRRSV antibody were detected in oral fluid samples by ELISA and IFAT. Although the approach remains to be validated in the field, the results of this experiment suggest that pen-based oral fluid sampling could be an efficient, costeffective approach to PRRSV surveillance in swine populations.
The purpose of this review was to discuss the history of the development and implementation of oral fluid diagnostics for infectious diseases of humans and domestic animals. The use of oral fluid for the assessment of health and diagnosis of disease in humans and animals has a surprisingly long history. As early as 1909, Pollaci and Ceraulo reported sensitive and specific agglutination of 'Micrococcus melitensis' (Brucella melitensis) by oral fluid from patients diagnosed with Malta Fever. A 1986 report of the detection of antibodies against human immunodeficiency virus (HIV) in oral fluid from patients with acquired immunodeficiency syndrome (AIDS) marked the start of a remarkably rapid series of developments in oral fluid-based assays. Cumulatively, the literature strongly supports implementation of oral fluid-based diagnostics in veterinary diagnostic medicine. Pathogen-specific IgA, IgM and IgG antibodies have all been demonstrated in oral fluid collected from diverse domestic animal species in response to infection. A variety of infectious agents, both local and systemic, are shed in oral fluid, including some of the most economically significant pathogens of production animals (e.g. foot-and-mouth disease virus, classical swine fever virus and porcine reproductive and respiratory syndrome virus) Ultimately, point-of-care rapid assays (i.e. cow-side, sow-side or pen-side tests) and access to real-time infectious disease data will revolutionize our delivery of health management services.
The purpose of the present study was to evaluate the diagnostic performance of a commercial serum antibody enzyme-linked immunosorbent assay (ELISA) modified to detect anti– Porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in pen-based oral fluid specimens. Experimental and field oral fluid samples of defined status in reference to exposure of swine with PRRSV were used to derive the kinetics of detectable concentrations of antibody against PRRSV. Immunoglobulin (Ig)M and IgA were readily detected in oral fluid specimens from populations in which PRRSV infection was synchronized among all individuals but not in samples collected in commecial herds. In contrast, IgG was readily detected at diagnostically useful levels in both experimental and field samples for up to 126 days. Estimates of the IgG oral fluid ELISA performance were based on results from testing positive oral fluid samples ( n = 492) from experimentally inoculated pigs ( n = 251) and field samples ( n = 241) and negative oral fluid samples ( n = 367) from experimentally inoculated pigs ( n = 84) and field samples ( n = 283). Receiver operating characteristic analysis estimated the diagnostic sensitivity and specificity of the assay as 94.7% (95% confidence interval [CI]: 92.4, 96.5) and 100% (95% CI: 99.0, 100.0), respectively, at a sample-to-positive ratio cutoff of ≥0.40. The results of the study suggest that the IgG oral fluid ELISA can provide efficient, cost-effective PRRSV monitoring in commercial herds and PRRSV surveillance in elimination programs.
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