Jin Li, Thomas Brenda
Virus Reference Department, Centre for Infections, Health Protection Agency, London, UK.
J Med Virol. 2007 Jul;79(7):1017-24. doi: 10.1002/jmv.20847.
Rubella and congenital rubella syndrome continue to be important health problems worldwide. The detection of rubella RNA directly in clinical specimens is a critical factor in early laboratory diagnosis of recent or congenital infection, in addition to detection of rubella-specific IgM. In order to comply with recent WHO recommendations for establishing uniform genetic analysis protocols for rubella virus we have developed a new block based PCR assay (PCR-E317), which extends the sequence generated by the block based PCR-E592 currently in use, to cover the minimum acceptable 739 nucleotides (nt) window at the E1 gene. In addition, a real-time PCR assay has been developed to allow rapid detection of the virus in the laboratory. The assays were applied to a number of clinical specimens collected from patients including recent rubella incidences in the UK, Ethiopia and Turkey, two prenatal and two congenital rubella syndrome cases. Rubella RNA was detected in specimens from two patients that were collected too early for IgM detection, in two amniotic fluids for prenatal diagnosis and in the follow up specimens from the two infant with congenital rubella syndrome tested for viral secretion. At least four genotypes were identified among these patients. The results showed that molecular assays are important tools in the early diagnosis of rubella and congenital rubella syndrome, in the provision of molecular epidemiological information for tracking transmission pathways and in adding to the knowledge of rubella strain distribution worldwide.
风疹和先天性风疹综合征仍是全球重要的健康问题。除检测风疹特异性IgM外，直接在临床标本中检测风疹RNA是近期感染或先天性感染早期实验室诊断的关键因素。为了遵循世界卫生组织（WHO）近期制定的风疹病毒统一基因分析方案的建议，我们开发了一种新的基于阻断的PCR检测方法（PCR-E317），该方法扩展了目前使用的基于阻断的PCR-E592所产生的序列，以覆盖E1基因至少739个核苷酸（nt）的可接受窗口。此外，还开发了一种实时PCR检测方法，以便在实验室中快速检测病毒。这些检测方法应用于从患者收集的一些临床标本，包括英国、埃塞俄比亚和土耳其近期的风疹病例，两例产前病例和两例先天性风疹综合征病例。在两名患者的标本中检测到风疹RNA，这些标本采集时间过早，无法检测到IgM；在两份用于产前诊断的羊水中以及在两名接受病毒分泌检测的先天性风疹综合征婴儿的随访标本中也检测到了风疹RNA。在这些患者中至少鉴定出四种基因型。结果表明，分子检测方法是风疹和先天性风疹综合征早期诊断的重要工具，可为追踪传播途径提供分子流行病学信息，并有助于增加对全球风疹毒株分布的了解。
