Oxford University Clinical Research Unit, Wellcome Trust Major Overseas Program, Ho Chi Minh City, Viet Nam.
J Virol Methods. 2012 Jan;179(1):250-5. doi: 10.1016/j.jviromet.2011.11.012. Epub 2011 Nov 19.
Improved diagnostic tools for rapid detection, quantitation, and subgrouping of human respiratory syncytial virus (RSV) are needed to aid the development and evaluation of novel intervention strategies. A quantitative real-time RT-PCR using specific locked nucleic acid (LNA) probes was developed to identify RSV and to distinguish RSV subgroups A and B (RSV LNA assay). RSV subgroup diversity and the relationship between viral load and disease severity in confirmed RSV infections were also explored. 264 archived respiratory specimens from pediatric patients were tested in parallel using the commercial multiplex Seeplex™ RV detection kit (Seegene) and the novel RSV LNA assay. The LNA assay demonstrated a significantly higher sensitivity than Seeplex, improving overall detection rates from 24% (64/264) to 32% (84/264). Detection limits of 9.0×10(1) and 6.0×10(2)copies/mL were observed for RSV A and B, respectively. RSV A was detected in 53/84 (63%) cases, and 31/84 (37%) were positive for RSV B. This novel method offers a rapid, quantitative, highly specific and sensitive approach to laboratory diagnosis of RSV.
需要改进的快速检测、定量和亚群人类呼吸道合胞病毒（RSV）的诊断工具，以帮助发展和评估新的干预策略。一种使用特定的锁核酸（LNA）探针的定量实时 RT-PCR 被开发出来，以鉴定 RSV 并区分 RSV 亚组 A 和 B（RSV LNA 检测）。还探讨了 RSV 亚群多样性以及病毒载量与确诊 RSV 感染严重程度之间的关系。使用商业多重 Seeplex ™ RV 检测试剂盒（Seegene）和新型 RSV LNA 检测试剂盒，对 264 份来自儿科患者的存档呼吸道标本进行平行检测。LNA 检测的灵敏度明显高于 Seeplex，总体检出率从 24%（64/264）提高到 32%（84/264）。分别观察到 RSV A 和 B 的检测限为 9.0×10(1)和 6.0×10(2)拷贝/ml。在 84 例（63%）中检测到 RSV A，31 例（37%）为 RSV B 阳性。这种新方法提供了一种快速、定量、高度特异和敏感的 RSV 实验室诊断方法。
