Shimizu C, Shimizu H, Mitsuda T, Tsukuda M, Ichikawa S, Yokota S
Department of Pediatrics, Yokohama City University School of Medicine, Kanagawa, Japan.
Mol Cell Probes. 1994 Jun;8(3):193-8. doi: 10.1006/mcpr.1994.1026.
A rapid and sensitive one-step polymerase chain reaction (PCR) assay was developed for use in identifying type I and type II herpes simplex virus (HSV). Although the nucleotide sequences of the two HSV subtypes are quite similar, common and type-specific sequences 20 nucleotides in length could be deduced in the thymidine kinase gene. Oligonucleotide primers targeted to the type-specific regions generated products of different sizes that served to distinguish two HSV types. Type-specific PCR amplification products were verified by restriction enzyme digestion. Specificity of the HSV PCR was established by the lack of amplification of other herpes-group viruses including cytomegalovirus, Epstein-Barr virus, and varicella zoster virus. Extraction of DNA from clinical materials (throat swabs, vesicular swabs, cerebrospinal fluid and eye discharge) yielded an amplification product of the predicted size for each HSV type. Thus, this PCR system provides a rapid, sensitive and specific assay that can supplement the currently available modalities for detecting and typing HSV.
开发了一种快速灵敏的一步法聚合酶链反应（PCR）检测方法，用于鉴定I型和II型单纯疱疹病毒（HSV）。尽管两种HSV亚型的核苷酸序列非常相似，但在胸苷激酶基因中可以推导得出长度为20个核苷酸的共同序列和型特异性序列。靶向型特异性区域的寡核苷酸引物产生了不同大小的产物，用于区分两种HSV类型。通过限制性酶切验证型特异性PCR扩增产物。HSV PCR的特异性通过未扩增包括巨细胞病毒、EB病毒和水痘带状疱疹病毒在内的其他疱疹病毒群病毒得以确立。从临床材料（咽拭子、水疱拭子、脑脊液和眼分泌物）中提取DNA，得到了每种HSV类型预测大小的扩增产物。因此，该PCR系统提供了一种快速、灵敏且特异的检测方法，可补充目前用于检测和分型HSV的现有方法。
