Nicoll S, Brass A, Cubie H A
Regional Clinical Virology Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland, UK.
J Virol Methods. 2001 Jul;96(1):25-31. doi: 10.1016/s0166-0934(01)00315-9.
Herpes viruses including cytomegalovirus, varicella zoster and herpes simplex are an important cause of morbidity and mortality, especially in immunocompromised patients. Real-time PCR assays were developed with the aim of introducing a rapid and sensitive test to replace culture, and as a surveillance system for high-risk patients. The assays were optimised using cell culture derived material, and the sensitivity ascertained using cloned product before applying to extracted and non-extracted clinical samples. The sensitivity was between 1--100 virus copies with increased sensitivity to detect less than 10 copies possible when an initial round of amplification was carried out using external primers. Results were available within four hours of receipt compared with a median of 4.4 days for culture and immunofluorescence. Real-time PCR was found to be a sensitive and rapid method of detecting these viruses and will be a valuable tool for the surveillance of immunosuppressed patients.
包括巨细胞病毒、水痘带状疱疹病毒和单纯疱疹病毒在内的疱疹病毒是发病和死亡的重要原因，尤其是在免疫功能低下的患者中。开发实时聚合酶链反应（PCR）检测方法的目的是引入一种快速、灵敏的检测方法来取代培养法，并作为高危患者的监测系统。使用细胞培养衍生材料对检测方法进行了优化，并在应用于提取和未提取的临床样本之前，使用克隆产物确定了灵敏度。灵敏度在1至100个病毒拷贝之间，当使用外部引物进行第一轮扩增时，检测少于10个拷贝的灵敏度可能会提高。与培养法和免疫荧光法的中位数4.4天相比，收到样本后4小时内即可获得结果。实时PCR被发现是检测这些病毒的一种灵敏且快速的方法，将成为监测免疫抑制患者的有价值工具。
