Carvalho R, Oliveira A M, Souza A M, Passos L M, Martins A S
Ministério da Agricultura e do Abastecimento, Belo Horizonte, MG, Brazil.
Arch Virol. 2000;145(9):1773-87. doi: 10.1007/s007050070055.
In this study, an improved polymerase chain reaction (PCR) was used for detection of DNA of latent EHV-1 strains from several sources. Three pairs of oligonucleotide primers spanning fragments of 333 bp, 226 bp and 268 bp of the thymidine kinase (tk) gene, and one primer pair spanning 225 bp of the glycoprotein C (gC) gene were used in specific amplifications. Primers for EHV-4 PCR were also designed. Restriction digests with TaqI confirmed the identity of tk PCR fragments from EHV-1. The sensitivity to detect PCR products was further improved by visualisation in silver-stained acrylamide gels. PCR assays were applied to 267 samples including pools of tissue, peripheral blood leukocytes (PBL) and nasal swabs of archived, farms and abattoir specimens from a total of 116 animals. The EHV-1 DNA was found in 88% of the analysed samples. The prevalence of the EHV-1 latent or persistent form in adult horses was similar to others reports but found higher than previously described in foetuses and young foals. EHV-4 latency was not detected in the Brazilian studied specimens.
在本研究中，采用改良的聚合酶链反应（PCR）检测来自多个来源的潜伏性马疱疹病毒1型（EHV-1）毒株的DNA。使用了三对跨越胸苷激酶（tk）基因333 bp、226 bp和268 bp片段的寡核苷酸引物，以及一对跨越糖蛋白C（gC）基因225 bp的引物进行特异性扩增。还设计了用于EHV-4 PCR的引物。用TaqI进行限制性酶切证实了来自EHV-1的tk PCR片段的同一性。通过银染聚丙烯酰胺凝胶可视化进一步提高了检测PCR产物的灵敏度。PCR检测应用于267个样本，包括来自总共116只动物的存档组织、外周血白细胞（PBL）和鼻拭子、农场和屠宰场标本的混合样本。在88%的分析样本中发现了EHV-1 DNA。成年马中EHV-1潜伏或持续感染形式的流行率与其他报告相似，但高于先前在胎儿和幼驹中的描述。在巴西研究的标本中未检测到EHV-4潜伏感染。
