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首页> 外文期刊>The journal of microbiology >Molecular Survey of Latent Pseudorabies Virus Infection in Nervous Tissues of Slaughtered Pigs by Nested and Real-time PCR
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Molecular Survey of Latent Pseudorabies Virus Infection in Nervous Tissues of Slaughtered Pigs by Nested and Real-time PCR

机译:巢式和实时荧光定量PCR技术对屠宰猪神经组织中潜伏性伪狂犬病病毒感染的分子生物学研究

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In this study, the prevalence and quantity of a latent pseudorabies virus (PrV) infection in the nervous tissues of randomly selected pigs was determined via nested and real-time PCR. The nervous tissues, including the trigeminal ganglion (TG), olfactory bulb (OB), and brain stem (BS), were collected from the heads of 40 randomly selected pigs. The majority of the nervous tissues from the selected pigs evidenced a positively amplified band on nested PCR. In particular, nested PCR targeted to the PrV glycoprotein B (gB) gene yielded positive results in all of the BS samples. Nested PCR for either the gE or gG gene produced positive bands in a less number of nervous tissues (57.5% and 42.5%, respectively). Real-time PCR revealed that the examined tissues harbored large copy numbers of latent PrV DNA, ranging between 100.1 and 107.2 (1-1.58x107) copies per 1 g of genomic DNA. Real-time PCR targeted to the PrV gE gene exhibited an accumulated fluorescence of reporter dye at levels above threshold, thereby indicating a higher prevalence than was observed on the nested PCR (100% for BS, 92% for OB, and 85% for TG). These results indicate that a large number of farm-grown pigs are latently infected with a field PrV strain with a variety of copy numbers. This result is similar to what was found in association with the human herpes virus.
机译:在这项研究中,通过巢式和实时PCR确定了随机选择的猪的神经组织中潜在的伪狂犬病病毒(PrV)感染的发生率和数量。从40只随机选择的猪的头部收集神经组织,包括三叉神经节(TG),嗅球(OB)和脑干(BS)。选定猪的大多数神经组织在巢式PCR上显示出一条阳性扩增带。特别是,靶向PrV糖蛋白B(gB)基因的巢式PCR在所有BS样品中均产生阳性结果。 gE或gG基因的巢式PCR在较少数量的神经组织中产生阳性条带(分别为57.5%和42.5%)。实时PCR显示,被检查的组织内藏有大量拷贝的潜伏PrV DNA,每1 g基因组DNA拷贝在100.1至107.2(1-1.58x107)个拷贝之间。靶向PrV gE基因的实时PCR在高于阈值的水平上表现出报告染料的累积荧光,因此表明其患病率高于巢式PCR(BS为100%,OB为92%,TG为85%) )。这些结果表明,大量的猪种潜伏地感染了具有多种拷贝数的野猪PrV株。该结果类似于与人疱疹病毒有关的发现。

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