Oxford University Clinical Research Unit, Hospital for Tropical Diseases, 190 Ben Ham Tu, Quan 5, Ho Chi Minh City, Viet Nam.
J Virol Methods. 2011 Nov;177(2):168-73. doi: 10.1016/j.jviromet.2011.08.002. Epub 2011 Aug 5.
Dengue is mosquito-borne virus infection that annually causes ~50 million clinically apparent cases worldwide. An internally controlled one-step real-time multiplex RT-PCR assay was developed for detection and quantitation of DENV RNA in plasma sample by using specific primers and fluorogenic TaqMan probes. All primers and probes targeted sequences near the 3' end of the NS5 gene. The method comprised two multiplex assays and was validated for sensitivity, specificity, linearity, reproducibility and precision. An internal control template was spiked into each clinical specimen to provide quality assurance for each experimental step. The assay allowed for detection of between 0.5 and 3 infectious particles per mL, is rapid and has been operationally characterized in 287 Vietnamese dengue patients from two therapeutic intervention trials at the Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam.
登革热是一种由蚊子传播的病毒感染，每年在全球范围内导致约 5000 万例临床明显病例。本研究开发了一种内部对照的一步实时多重 RT-PCR 检测法，使用特异性引物和荧光 TaqMan 探针，对血浆样本中的 DENV RNA 进行检测和定量。所有引物和探针均针对 NS5 基因 3'端附近的序列。该方法包括两个多重检测，对其灵敏度、特异性、线性、重复性和精密度进行了验证。在每份临床标本中加入内部对照模板，以保证每个实验步骤的质量。该检测方法可检测到 0.5 至 3 个感染性颗粒/毫升，快速且已在越南胡志明市热带病医院的两项治疗干预试验中的 287 例越南登革热患者中进行了操作特征描述。
