TY - JOUR
T1 - Evaluation of the sensitivity of reverse-transcription polymerase chain reaction to detect porcine reproductive and respiratory syndrome virus on individual and pooled samples from boars
AU - Rovira, Albert
AU - Clement, Travis
AU - Christopher-Hennings, Jane
AU - Thompson, Bob
AU - Engle, Mark
AU - Reicks, Darwin
AU - Muñoz-Zanzi, Claudia
PY - 2007/9
Y1 - 2007/9
N2 - Boar studs are continuously monitored for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) by testing different biological samples by reverse-transcription polymerase chain reaction (RT-PCR). In most cases, samples are run in pools, even though the impact of pooling on the sensitivity of RT-PCR is unknown. The objective of this study was to evaluate the feasibility of using PCR on pooled samples through the estimation of the sensitivity of RT-PCR on different biological samples run individually, in pools of 3 and in pools of 5. Twenty-nine boars were inoculated with a low virulent PRRSV isolate. Serum, blood swab, and semen samples were obtained from each boar every 2 to 3 days for 2 weeks. Each sample was tested by RT-PCR undiluted or diluted 1:3 and 1:5 with negative samples. Eleven of the 29 boars did not appear to get infected from the inoculum, as evidenced by no seroconversion 15 days after inoculation. Data from the other 18 boars showed that serum was the best sample to detect PRRSV during acute infection, with the blood swab sample performing almost as well. Semen samples failed to detect PRRSV infection in most of the cases. Pooling samples at pool sizes of 3 and 5 resulted in a decrease in the sensitivity of RT-PCR. Sensitivity was reduced by 6% and 8%, respectively, when serum or blood swab samples were run in pools of 5. The impact of pooling on the sensitivity of PCR was higher in samples taken during the beginning of the viremic period.
AB - Boar studs are continuously monitored for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) by testing different biological samples by reverse-transcription polymerase chain reaction (RT-PCR). In most cases, samples are run in pools, even though the impact of pooling on the sensitivity of RT-PCR is unknown. The objective of this study was to evaluate the feasibility of using PCR on pooled samples through the estimation of the sensitivity of RT-PCR on different biological samples run individually, in pools of 3 and in pools of 5. Twenty-nine boars were inoculated with a low virulent PRRSV isolate. Serum, blood swab, and semen samples were obtained from each boar every 2 to 3 days for 2 weeks. Each sample was tested by RT-PCR undiluted or diluted 1:3 and 1:5 with negative samples. Eleven of the 29 boars did not appear to get infected from the inoculum, as evidenced by no seroconversion 15 days after inoculation. Data from the other 18 boars showed that serum was the best sample to detect PRRSV during acute infection, with the blood swab sample performing almost as well. Semen samples failed to detect PRRSV infection in most of the cases. Pooling samples at pool sizes of 3 and 5 resulted in a decrease in the sensitivity of RT-PCR. Sensitivity was reduced by 6% and 8%, respectively, when serum or blood swab samples were run in pools of 5. The impact of pooling on the sensitivity of PCR was higher in samples taken during the beginning of the viremic period.
KW - Blood swab
KW - Boar
KW - PCR
KW - Pooling
KW - Prrs virus
KW - Semen
KW - Sensitivity
KW - Serum
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UR - http://www.scopus.com/inward/citedby.url?scp=38449119381&partnerID=8YFLogxK
U2 - 10.1177/104063870701900507
DO - 10.1177/104063870701900507
M3 - Article
C2 - 17823393
AN - SCOPUS:38449119381
SN - 1040-6387
VL - 19
SP - 502
EP - 509
JO - Journal of Veterinary Diagnostic Investigation
JF - Journal of Veterinary Diagnostic Investigation
IS - 5
ER -