Fedele C G, Negredo A, Molero F, Sánchez-Seco M P, Tenorio A
Alerts and Emergencies Unit, National Center of Microbiology, Instituto de Salud Carlos III, Ctra. Pozuelo Majadahonda Km 2, 28220 Majadahonda, Madrid, Spain.
J Clin Microbiol. 2006 Dec;44(12):4464-70. doi: 10.1128/JCM.00276-06. Epub 2006 Oct 25.
Smallpox, once a devastating disease caused by Variola virus, a member of the Orthopoxvirus genus, was eradicated in 1980. However, the importance of variola virus infections has been stressed widely in the last few years, particularly following recent social events in the world. Today, variola virus is considered to be one of the most significant agents with potential use as a biological weapon. In this study we developed an internally controlled real-time PCR assay for rapid detection and simultaneous differentiation of variola virus from other orthopoxviruses. The assay is based on TaqMan 3'-minor groove binder (MGB) chemistry and uses generic primers, designed in highly conserved genomic regions of the crmB gene, and three TaqMan MGB probes designed to identify orthopoxviruses, variola virus, and an internal control. The results obtained suggest that the assay is rapid, sensitive, specific, and suitable for the generic detection of orthopoxviruses and the identification of variola virus and avoids false-negative results in a single reaction tube.
天花曾是一种由正痘病毒属成员天花病毒引起的毁灭性疾病，于1980年被根除。然而，在过去几年里，天花病毒感染的重要性受到了广泛强调，特别是在最近世界上发生的一些社会事件之后。如今，天花病毒被认为是最有可能被用作生物武器的重要病原体之一。在本研究中，我们开发了一种内部对照实时PCR检测方法，用于快速检测天花病毒并同时将其与其他正痘病毒区分开来。该检测方法基于TaqMan 3'-小沟结合剂（MGB）化学原理，使用在crmB基因高度保守的基因组区域设计的通用引物，以及设计用于鉴定正痘病毒、天花病毒和一个内部对照的三种TaqMan MGB探针。获得的结果表明，该检测方法快速、灵敏、特异，适用于正痘病毒的通用检测以及天花病毒的鉴定，并且避免了在单个反应管中出现假阴性结果。
