Harbecke Ruth, Oxman Michael N, Arnold Beth A, Ip Charlotte, Johnson Gary R, Levin Myron J, Gelb Lawrence D, Schmader Kenneth E, Straus Stephen E, Wang Hui, Wright Peter F, Pachucki Constance T, Gershon Anne A, Arbeit Robert D, Davis Larry E, Simberkoff Michael S, Weinberg Adriana, Williams Heather M, Cheney Carol, Petrukhin Luba, Abraham Katalin G, Shaw Alan, Manoff Susan, Antonello Joseph M, Green Tina, Wang Yue, Tan Charles, Keller Paul M
Department of Veterans Affairs San Diego Healthcare System, San Diego, California, USA.
J Med Virol. 2009 Jul;81(7):1310-22. doi: 10.1002/jmv.21506.
A real-time PCR assay was developed to identify varicella-zoster virus (VZV) and herpes simplex virus (HSV) DNA in clinical specimens from subjects with suspected herpes zoster (HZ; shingles). Three sets of primers and probes were used in separate PCR reactions to detect and discriminate among wild-type VZV (VZV-WT), Oka vaccine strain VZV (VZV-Oka), and HSV DNA, and the reaction for each virus DNA was multiplexed with primers and probe specific for the human beta-globin gene to assess specimen adequacy. Discrimination of all VZV-WT strains, including Japanese isolates and the Oka parent strain, from VZV-Oka was based upon a single nucleotide polymorphism at position 106262 in ORF 62, resulting in preferential amplification by the homologous primer pair. The assay was highly sensitive and specific for the target virus DNA, and no cross-reactions were detected with any other infectious agent. With the PCR assay as the gold standard, the sensitivity of virus culture was 53% for VZV and 77% for HSV. There was 92% agreement between the clinical diagnosis of HZ by the Clinical Evaluation Committee and the PCR assay results.
开发了一种实时聚合酶链反应（PCR）检测方法，用于鉴定疑似带状疱疹（HZ；缠腰火丹）患者临床标本中的水痘-带状疱疹病毒（VZV）和单纯疱疹病毒（HSV）DNA。在单独的PCR反应中使用了三组引物和探针，以检测和区分野生型VZV（VZV-WT）、Oka疫苗株VZV（VZV-Oka）和HSV DNA，并且针对每种病毒DNA的反应与针对人β-珠蛋白基因的特异性引物和探针进行多重反应，以评估标本的充足性。基于开放阅读框62中第106262位的单核苷酸多态性，将所有VZV-WT毒株（包括日本分离株和Oka亲本株）与VZV-Oka区分开来，从而导致同源引物对的优先扩增。该检测方法对目标病毒DNA具有高度敏感性和特异性，未检测到与任何其他感染因子的交叉反应。以PCR检测作为金标准，病毒培养对VZV的敏感性为53%，对HSV的敏感性为77%。临床评估委员会对HZ的临床诊断与PCR检测结果之间的一致性为92%。
