Katz Russell S, Premenko-Lanier Mary, McChesney Michael B, Rota Paul A, Bellini William J
Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Med Virol. 2002 Aug;67(4):596-602. doi: 10.1002/jmv.10144.
The purpose of this study was to evaluate the use of dried blood spots stored on filter paper as a means to provide specimens for virologic surveillance for measles virus (MV) in situations when the reverse cold chain is not available. Two single-step RT-PCR assays were evaluated for sensitivity of detection of MV nucleoprotein gene RNA. The more sensitive assay was then used to assess the stability of MV RNA in dried whole blood stored on filter paper. MV RNA was found to be stable in dried blood spots for up to 2 months at room temperature or 1 month at 37 degrees C. As few as 100 infected human peripheral blood mononuclear cells (PBMC) per blood spot could be detected using a single-step RT-PCR reaction and ethidium bromide detection. MV RNA was also detected in dried blood spots obtained from rhesus macaques after challenge with wild-type MV. In the rhesus samples, the single-step RT-PCR reaction could detect approximately 10(3) TCID(50) per blood spot, while nested PCR detected 3 TCID(50) per blood spot. The results of this laboratory-based study suggest that the use of dried blood spots stored on filter has the potential to improve virologic surveillance for MV in some areas, and they emphasize the need for continued testing under field conditions.
本研究的目的是评估在没有反向冷链的情况下，使用滤纸保存的干血斑作为提供麻疹病毒（MV）病毒学监测标本的一种手段。对两种单步逆转录聚合酶链反应（RT-PCR）检测方法检测MV核蛋白基因RNA的敏感性进行了评估。然后使用更敏感的检测方法评估MV RNA在滤纸保存的干全血中的稳定性。发现MV RNA在干血斑中于室温下可稳定保存长达2个月，在37℃下可稳定保存1个月。使用单步RT-PCR反应和溴化乙锭检测，每个血斑中低至100个受感染的人外周血单个核细胞（PBMC）都可被检测到。在用野生型MV攻击恒河猴后获得的干血斑中也检测到了MV RNA。在恒河猴样本中，单步RT-PCR反应每个血斑可检测到约10(3)半数组织培养感染剂量（TCID(50)），而巢式PCR每个血斑可检测到3个TCID(50)。这项基于实验室的研究结果表明，使用滤纸保存的干血斑有可能改善某些地区对MV的病毒学监测，并且强调了在现场条件下继续进行检测的必要性。
