Yoon Hyun A, Eo Seong Kug, Aleyas Abi George, Park Seong Ok, Lee John Hwa, Chae Joon Seok, Cho Jeong Gon, Song Hee Jong
Laboratory of Microbiology, College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University, Jeonju 561-756, Republic of Korea.
J Microbiol. 2005 Oct;43(5):430-6.
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 10(0.1) and 10(7.2) (1-1.58 x 10(7)) copies per 1 microg 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.
在本研究中,通过巢式PCR和实时PCR测定了随机选取猪的神经组织中潜伏性伪狂犬病病毒(PrV)感染的患病率和病毒量。从40头随机选取猪的头部采集神经组织,包括三叉神经节(TG)、嗅球(OB)和脑干(BS)。所选猪的大多数神经组织在巢式PCR上显示出阳性扩增条带。特别是,针对PrV糖蛋白B(gB)基因的巢式PCR在所有BS样本中均产生阳性结果。针对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,所检测的组织中含有大量潜伏性PrV DNA拷贝数,每1微克基因组DNA介于10(0.1)至10(7.2)(1 - 1.58×10(7))拷贝之间。针对PrV gE基因的实时PCR显示报告染料的累积荧光高于阈值水平,从而表明患病率高于巢式PCR所观察到的结果(BS为100%,OB为92%,TG为85%)。这些结果表明,大量养殖猪被具有多种拷贝数的田间PrV毒株潜伏感染。这一结果与人类疱疹病毒的相关发现相似。