| 权娅茹,陈震,邱平,崔晓雨,李长贵,袁力勇.水痘带状疱疹病毒野毒株和疫苗株区分方法研究[J].中国药事,2017,(9):1026-1031 |
| 水痘带状疱疹病毒野毒株和疫苗株区分方法研究 |
| On the Method of Distinguishing Varicella-zoster Virus Wild-type Strains from Vaccine Strains |
| 投稿时间：2017-02-22 |
| DOI：10.16153/j.1002-7777.2017.09.012 |
| 中文关键词: 水痘带状疱疹病毒 野毒株 疫苗株 ORF62区 PCR-RFLP |
| 英文关键词: varicella-zoster virus wild-type strain vaccine strain ORF62 region PCR-RFLP |
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| 中文摘要: |
| 目的：建立水痘带状疱疹病毒野毒株和疫苗株的聚合酶链反应和限制性片段多态性（PCRRFLP）区分方法。方法：对疫苗株和水痘临床患者疱疹液进行开放阅读框62（ORF62）区基因序列测定，确定可用于野毒株和疫苗株区分的单核苷酸的多态性（SNP）位点。PCR扩增含SmaⅠ、BssHⅡ和NaeⅠ酶切位点的基因（DNA）片段后进行限制性内切酶酶切，利用酶切图谱区分野毒株和疫苗株。结果：用PCR-RFLP分析疫苗株和疱疹液，疫苗株酶切图谱为SmaⅠ+BssHⅡ+NaeⅠ+，疱疹液酶切图谱为SmaⅠ-BssHⅡ-NaeⅠ-。结论：基于ORF62区的106262（SmaⅠ）、107136（BssHⅡ）、107252（NaeⅠ）的PCR-RFLP可有效区分野毒株和疫苗株。 |
| 英文摘要: |
| Objective:To establish a method to distinguish varicella-zoster virus wild-type strain from vaccine strain by using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). Methods:The open reading frame 62 (ORF62) genes of vaccine strain and herpes fluid of inclinical patients with chickenpox were sequenced to determine the SNP location which could be used to distinguish the wild-type strain from the vaccine strain. DNA fragments containing sites of restriction endonucleases of SmaⅠ, BssHⅡ, and NaeⅠ were amplified by PCR, and then the PCR products were digested by the restriction endonucleases. Finally, the RFLPs generated by restriction digestion could be used to distinguish the wild-type strain from the vaccine strain. Results:Vaccine strain and herpes fluid were analyzed using PCR-RFLP, vaccine strain was revealed by the triple positive of SmaⅠ+ BssHⅡ+ NaeⅠ+, herpes fluid was revealed by the triple negative of SmaⅠ- BssHⅡ- NaeⅠ-. Conclusion:A PCR-RFLP based on the 106262(SmaⅠ), 107136(BssHⅡ), and 107252(NaeⅠ) within the ORF62 region was effective in distinguishing the wild-type strain from the vaccine strain. |
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