Division of Viral Diseases, National Center for Immunizations and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, United States.
J Virol Methods. 2013 Feb;187(2):284-7. doi: 10.1016/j.jviromet.2012.11.023. Epub 2012 Nov 29.
Information on the molecular epidemiology of rubella has been valuable in supporting efforts to control and eliminate rubella in several countries. The preferred samples for virus isolation or RNA detection, such as throat swabs, are often not available making it difficult to obtain a robust database of rubella virus sequences. A method for obtaining rubella virus genotypes from more commonly collected samples such as sera or dried blood spots using real-time RT-PCR to screen samples followed by nested set amplification is described. Rubella genotypes were obtained from dried blood spots and recent and archival sera collections. Eighteen percent of the RNAs extracted from the archival sera were real-time RT-PCR positive, and 44% of these RNAs were amplified successfully by nested RT-PCR and sequenced. Implementation of this technique could provide another tool to improve global rubella molecular surveillance.
有关风疹分子流行病学的信息对于支持几个国家控制和消除风疹的努力非常有价值。病毒分离或 RNA 检测的首选样本，如咽喉拭子，通常不可用，因此难以获得风疹病毒序列的强大数据库。本文描述了一种使用实时 RT-PCR 对样本进行筛选，然后进行巢式套式扩增，从更常见采集的样本（如血清或干血斑）中获取风疹病毒基因型的方法。从干血斑和近期及存档血清采集物中获得了风疹病毒基因型。从存档血清中提取的 RNA 中，有 18%经实时 RT-PCR 检测为阳性，其中 44%通过巢式 RT-PCR 扩增成功并测序。该技术的实施可以为改善全球风疹分子监测提供另一种工具。
