Madsen E S, Madsen K G, Nielsen J, Jensen M H, Lei J C, Have P
Danish Veterinary Institute For Virus Research, Kalvehave, Denmark.
Vet Microbiol. 1997 Jan;54(1):1-16. doi: 10.1016/s0378-1135(96)01270-9.
The humoral antibody response against the nonstructural protein NS1 and the structural protein VP2 of porcine parvovirus (PPV) was evaluated by immuno-peroxidase test (IPT) and enzyme linked immuno sorbent assay (ELISA) using recombinant PPV antigens. The coding sequence for NS1 and VP2 was inserted into the baculovirus. Autographa californica nuclear polyhedrosis virus (AcNPV) genome resulting in two recombinant baculoviruses AcNPV-NS1 and AcNPV-VP2, respectively. Sf9 cells (Spodoptora frugidiperda) inoculated with AcNPV-NS1 producing recombinant nonstructural protein (rNS1) and AcNPV-VP2 producing recombinant virion protein (rVP2) were used in IPT and ELISA to analyse serum antibodies. Pigs vaccinated with an inactivated whole virus vaccine and experimentally infected pigs were studied. Significant titers against rVP2 were obtained in both vaccinated and infected pigs. Specific antibodies against rNS1 could only be detected in infected pigs and NS1 may in this way allow the specific detection of infected animals. Analysis of serum samples collected up to 18 days post infection (p.i.) from four pigs experimentally infected with PPV showed that antibodies against rNS1 and rVP2 could in all cases be detected on day 9 p.i. Two individual pigs were inoculated twice with PPV and the antibody response was followed 89 days after second inoculation. Serum antibodies against both rVP2 and rNS1 could be detected for this period of time.
使用重组猪细小病毒(PPV)抗原,通过免疫过氧化物酶试验(IPT)和酶联免疫吸附测定(ELISA)评估了针对PPV非结构蛋白NS1和结构蛋白VP2的体液抗体反应。将NS1和VP2的编码序列插入杆状病毒苜蓿银纹夜蛾核型多角体病毒(AcNPV)基因组中,分别产生两种重组杆状病毒AcNPV-NS1和AcNPV-VP2。用产生重组非结构蛋白(rNS1)的AcNPV-NS1和产生重组病毒粒子蛋白(rVP2)的AcNPV-VP2接种草地贪夜蛾细胞(Spodoptora frugidiperda),用于IPT和ELISA分析血清抗体。对接种灭活全病毒疫苗的猪和实验感染猪进行了研究。在接种疫苗和感染的猪中均获得了针对rVP2的显著滴度。仅在感染猪中检测到针对rNS1的特异性抗体,NS1可能以这种方式实现对感染动物的特异性检测。对4只实验感染PPV的猪在感染后(p.i.)长达18天收集的血清样本进行分析,结果显示在感染后第9天在所有情况下均可检测到针对rNS1和rVP2的抗体。两只猪用PPV接种两次,并在第二次接种后89天跟踪抗体反应。在此期间可检测到针对rVP2和rNS1的血清抗体。