Wills R W, Zimmerman J J, Yoon K J, Swenson S L, McGinley M J, Hill H T, Platt K B, Christopher-Hennings J, Nelson E A
Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Iowa State University, Ames 50011-1250, USA.
Vet Microbiol. 1997 Apr;55(1-4):231-40. doi: 10.1016/s0378-1135(96)01337-5.
Persistent infection with porcine reproductive and respiratory syndrome virus (PRRSV) was shown in experimentally infected pigs by isolation of virus from oropharyngeal samples for up to 157 days after challenge. Four 4 week old, conventional, PRRSV antibody-negative pigs were intranasally inoculated with PRRSV (ATCC VR-2402). Serum samples were collected every 2 to 3 days until day 42 post inoculation (PI), then approximately every 14 days until day 213 PI. Fecal samples were collected at the time of serum collection through day 35 PI. Oropharyngeal samples were collected at the time of serum collection from 56 to 213 days PI by scraping the oropharyngeal area with a sterile spoon, especially targeting the palatine tonsil. Turbinate, tonsil, lung, parotid salivary gland, spleen, lymph nodes and serum were collected postmortem on day 220 PI. Virus isolation (VI) on porcine alveolar macrophage cultures was attempted on all serum, fecal and oropharyngeal samples, as well as tissues collected postmortem. Postmortem tonsil tissues and selected fecal samples were also assayed for the presence of PRRSV RNA by the polymerase chain reaction (PCR). Serum antibody titers were determined by IFA, ELISA and SVN. Virus was isolated from all serum samples collected on days 2 to 11 PI and intermittently for up to 23 days in two pigs. No PRRSV was isolated from fecal samples, but 3 of 24 samples were PCR positive, suggesting the presence of inactivated virus. Oropharyngeal samples from each pig were VI positive 1 or more times between 56 and 157 days PI. Oropharyngeal samples from 3 of 4 pigs were VI positive on days 56, 70 and 84 PI. Virus was isolated from one pig on day 157 PI, 134 days after the last isolation of virus from serum from this animal. Virus was isolated from oropharyngeal samples for several weeks after the maximum serum antibody response, as measured by IFA, ELISA and SVN tests. All tissues collected postmortem were VI negative and postmortem tonsil samples were also negative by PCR. An important element in the transmission of PRRSV is the duration of virus shedding. The results of this study provided direct evidence of persistent PRRSV infection and explain field observations of long-term herd infection and transmission via purchase of clinically normal, but PRRSV infected, animals. Effective prevention and control strategies will need to be developed in the context of these results.
在实验感染的猪中,通过从口咽样本中分离病毒,显示猪繁殖与呼吸综合征病毒(PRRSV)持续感染长达攻毒后157天。将4头4周龄、常规的、PRRSV抗体阴性的猪经鼻接种PRRSV(ATCC VR - 2402)。每2至3天采集血清样本直至接种后42天(PI),然后大约每14天采集一次直至接种后213天PI。在采集血清样本时同时采集粪便样本直至接种后35天。在接种后56至213天采集血清样本时,用无菌匙刮取口咽区域,尤其针对腭扁桃体,采集口咽样本。在接种后220天对鼻甲、扁桃体、肺、腮腺、脾脏、淋巴结和血清进行尸检采集。对所有血清、粪便和口咽样本以及尸检采集的组织尝试在猪肺泡巨噬细胞培养物上进行病毒分离(VI)。对尸检扁桃体组织和选定的粪便样本也通过聚合酶链反应(PCR)检测PRRSV RNA的存在。通过间接荧光抗体试验(IFA)、酶联免疫吸附测定(ELISA)和病毒中和试验(SVN)测定血清抗体滴度。在接种后2至11天采集的所有血清样本中均分离到病毒,并且在两头猪中病毒间歇性分离长达23天。粪便样本中未分离到PRRSV,但24个样本中有3个PCR呈阳性,表明存在灭活病毒。每头猪的口咽样本在接种后56至157天之间有1次或更多次VI呈阳性。4头猪中有3头猪的口咽样本在接种后56、70和84天VI呈阳性。在接种后157天从一头猪中分离到病毒,这是该动物血清中最后一次分离到病毒后的134天。通过IFA、ELISA和SVN试验测定,在血清抗体反应达到最大值后的数周内,仍从口咽样本中分离到病毒。所有尸检采集的组织VI均为阴性,尸检扁桃体样本PCR也为阴性。PRRSV传播的一个重要因素是病毒排出的持续时间。本研究结果提供了PRRSV持续感染的直接证据,并解释了在现场观察到的长期群体感染以及通过购买临床正常但感染PRRSV的动物进行传播的现象。需要根据这些结果制定有效的预防和控制策略。