两种PCR -酶切酶切法检测皮肤粘膜或皮肤病变患者单纯疱疹病毒并分型的比较

Emma Herrera-Martínez, Rodolfo Ondarza-Aguilera, Sergio Estrada-Parra, Guillermo Pérez, Blanca L Barrón
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摘要

许多不同的聚合酶链反应(pcr)已经被描述用于检测和分型单纯疱疹病毒(HSV)。本文比较了两种酶切法(PCR/RFLP)检测和分型HSV的方法。设计了一套引物来扩增来自UL30和ul15基因的HSV DNA片段。分别用Ava II和Hpa II酶限制UL30和UL15扩增子进行分型。用两种参考菌株(HSV-1 McIntyre和HSV-2 G)和47株临床HSV分离株对该策略进行了测试。两种pcr都产生了预期的扩增子(一个492 bp的UL30和一个305 bp的UL15)。这两个扩增子的限制清楚地区分了HSV-和HSV-2,并产生了相同的结果。31株(66%)分离株为1型单纯疱疹病毒,16株(34%)分离株为2型单纯疱疹病毒。大多数HSV-1分离株(27/31)来自口面部和胸部病变;2型单纯疱疹病毒分离株中有1 / 2(8/16)来自同一解剖区域。结果表明,两种PCR/RFLP均可用于HSV的检测和分型。此外,我们对HSV-1和HSV-2感染解剖部位的研究结果与之前报道的疱疹病毒感染经典解剖定位的变化一致。
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Comparison of two methods of PCR followed by enzymatic restriction digestion for detection and typing of herpes simplex viruses isolated from patients with mucocutaneous or cutaneous lesions.

A multitude of different polymerase chain reactions (PCRs) have been described for detection and typing of Herpes simplex virus (HSV). This paper compares two PCRs coupled to enzymatic restriction (PCR/RFLP) to detect and type HSV. A primers set was designed to amplify a HSV DNA fragment from UL30 and UL 15 genes. Typing was done by restriction of the UL30 and UL15 amplicons with Ava II and Hpa II enzymes, respectively. This strategy was tested with two reference strains (HSV-1 McIntyre, and HSV-2 G), and 47 clinical HSV isolates. Both PCRs produced the expected amplicons (a 492 bp UL30, and 305 bp UL15). The restriction of both amplicons clearly differentiated HSV- from HSV-2, and produced equal results. Thirty one (66%) of the isolates were identified as HSV-1, and the other 16 (34%), as HSV-2. Most of the HSV-1 isolates (27/31) were from orofacial and thoracic lesions; and also, one half of the HSV-2 isolates (8/16) were from the same anatomical regions. Our results showed that either of the two PCR/RFLP could be used to detect and type HSV. Furthermore, our results of the anatomical site of HSV-1 and HSV-2 infections are consistent with previous reports which have shown changes in the classical anatomical localization of herpesvirus infections.

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