Comparison of Specimens for Detection of Porcine Reproductive and Respiratory Syndrome Virus Infection in Boar Studs
Comparison of Specimens for Detection of Porcine Reproductive and Respiratory Syndrome Virus Infection in Boar Studs
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DOI：
10.1111/tbed.12135
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发表时间：
2015-06-01
影响因子：
4.3
通讯作者：
Zimmerman, J.
中科院分区：
文献类型：
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作者：
Pepin, B. J.;Kittawornrat, A.;Zimmerman, J.
被猪繁殖与呼吸综合征病毒(PRRSV)污染的公猪精液是传播给雌性的途径，及早发现公猪中的PRRSV感染是母猪养殖场生物安全的关键组成部分。本研究的目的是确定检测猪急性PRRSV感染的最佳诊断标本(S)。对15头散养公猪进行精液和口腔液采集训练，然后接种商业PRRSV改良活病毒疫苗。从接种疫苗之日起至之后的14天内，每天采集所有公猪的口腔液样本。15头公猪被分成3组，每组5头，在精液采集时采集血清、血拭子和泡沫唾液，每3天一次。发泡的唾液来自下颌下唾液腺，是由被唤醒的公猪产生的。精液离心后，分别检测精液上清液和细胞组分。对所有样本进行随机排序，用PRRSV实时定量逆转录聚合酶链式反应(RRT-PCR)和抗体夹心法进行检测。在这项研究中，对血清、血拭子和口腔液的RRT-PCR结果进行了比较，发现在检测开始或阳性比例方面没有统计学意义上的差异，但在早期检测方面，血清在数量上优于口服液。免疫后第5天，血清和口服液的RRT-PCR结果一致。同样，在血清、口腔液和泡沫唾液中检测PRRSV抗体的开始时间在统计学上是相同的，血清结果再次显示出数字优势。这些结果表明，向母猪场提供PRRSV阴性精液的最高保证应该是对精液采集时采集的血清进行RRT-PCR检测。这种方法可以通过从随机选择的未收集的公猪中采集口服液样本来加强，以提供对公猪种种的统计有效监测。
Porcine reproductive and respiratory syndrome virus (PRRSV)-contaminated semen from boars is a route of transmission to females, and early detection of PRRSV infection in boars is a key component in sow farm biosecurity. The purpose of this study was to determine the optimum diagnostic specimen(s) for the detection of acute PRRSV infection in boars. Individually housed boars (n=15) were trained for semen and oral fluid collection and then vaccinated with a commercial PRRSV modified live virus vaccine. Starting on the day of vaccination and for 14days thereafter, oral fluid specimens were collected daily from all boars. The 15 boars were subdivided into three groups of 5, and serum, blood swabs and frothy saliva' were collected at the time of semen collection on a 3-day rotation. Frothy saliva, derived from the submandibular salivary gland, is produced by aroused boars. Semen was centrifuged, and semen supernatant and cell fractions were tested separately. All samples were randomly ordered and then tested by PRRSV real-time quantitative reverse-transcription polymerase chain reaction assay (rRT-PCR) and PRRSV antibody ELISA. In this study, a comparison of serum, blood swab, and oral fluid rRT-PCR results found no statistically significant differences in the onset of detection or proportion of positives, but serum was numerically superior to oral fluids for early detection. Serum and oral fluid provided identical rRT-PCR results at 5day post-vaccination. Likewise, the onset of detection of PRRSV antibody in serum, oral fluid and frothy saliva was statistically equivalent, with serum results again showing a numerical advantage. These results showed that the highest assurance of providing PRRSV-negative semen to sow farms should be based on rRT-PCR testing of serum collected at the time of semen collection. This approach can be augmented with oral fluid sampling from a random selection of uncollected boars to provide for statistically valid surveillance of the boar stud.