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Detection of challenge virus in fetal tissues by nested PCR as a test of the potency of a porcine parvovirus vaccine.

机译:巢式PCR检测胎儿组织中的攻击病毒作为猪节目病毒疫苗效力的试验。

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摘要

To estimate the potency of a porcine parvovirus (PPV) vaccine, 3 vaccinated and 3 non-vaccinated pregnant gilts were infected with PPV and the distribution of the virus was studied in the tissues of their 51 fetuses. Virus detection was attempted using haemagglutination (HA) and immunofluorescence (IF) assays, as well as by standard (single) and nested polymerase chain reactions (PCR). None of the detection methods yielded positive results when used to test for virus in suspensions of organs from the fetuses from the vaccinated gilts. The virus was detected in the fetuses from non-vaccinated gilts as follows: HA was positive in 14 of 23 (60.8%), IF in 16 of 23 (69.5%), standard PCR in 12 of 20 (60%), and the nested PCR in 19 of 23 (82.6%). Although the correlation among the results of various methods of virus detection was rather close (r < 0.83), the sensitivity of the nested PCR was the highest, both when testing dilutions of PPV and when examining the fetal organs. The nested PCR therefore provides a reliable approach for studies of virus distribution in fetal organs, with special reference to potency tests on vaccines.
机译:为了估算猪节目病毒(PPV)疫苗的效力,用PPV感染3疫苗和3个非接种疫苗的孕妇,并在其51胎儿的组织中研究了病毒的分布。使用血凝(HA)和免疫荧光(IF)测定以及标准(单)和巢式聚合酶链反应(PCR)来尝试病毒检测。当用于测试来自接种疫苗的胎儿的胎儿的胎儿的悬浮液中的病毒,没有任何检测方法产生阳性结果。从非接种疫苗的诱饵中检测到病毒,如下:如果在23(69.5%),21个(60%)的标准PCR中,则为23(60.8%)的14%(60.8%),HA为阳性。嵌套的PCR 19共分23(82.6%)。虽然各种病毒检测方法的结果之间的相关性相当接近(R <0.83),但是在测试PPV的稀释液和检查胎儿器官时,嵌套PCR的敏感性最高。因此,嵌套的PCR提供了一种可靠的方法,用于研究胎儿器官的病毒分布,特别是疫苗上的效力试验。

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