Borchers K, Slater J
Institut für Virologie der Freien Universität Berlin, Germany.
J Virol Methods. 1993 Dec 31;45(3):331-6. doi: 10.1016/0166-0934(93)90117-a.
The nested PCR method was applied for the detection and direct differentiation of equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4). Primer pairs were chosen from the glycoprotein B (gB) coding region of each serotype. The outer and inner EHV-1 primer pairs were type-specific, whereas the outer EHV-4 primer pair amplified EHV-1 and EHV-4 DNA and was therefore suitable for the detection of both virus types in a single sample. However, the nested EHV-4 primer pair was type-specific. The advantages of the nested PCR are twofold. Firstly, this assay was in the case of EHV-1 100 times, and in the case of EHV-4 1000 times more sensitive than the standard PCR, which indicates a detection limit of 1-0.1 fg of DNA (3-1 genome equivalents). Secondly, it allows the direct differentiation of EHV-1 and -4 without the need to resort to further analytical techniques.
巢式PCR方法用于检测和直接鉴别1型马疱疹病毒（EHV-1）和4型马疱疹病毒（EHV-4）。引物对选自各血清型的糖蛋白B（gB）编码区。EHV-1的外引物对和内引物对具有型特异性，而EHV-4的外引物对可扩增EHV-1和EHV-4的DNA，因此适用于在单个样本中检测这两种病毒类型。然而，巢式EHV-4引物对具有型特异性。巢式PCR的优点有两个。首先，该检测方法对于EHV-1而言比标准PCR敏感100倍，对于EHV-4而言比标准PCR敏感1000倍，这表明其DNA检测限为1-0.1 fg（3-1个基因组当量）。其次，它无需借助进一步的分析技术就能直接鉴别EHV-1和EHV-4。
