Prevention of Veterinary Medicine Department, College of Agriculture, Yanbian University, Yanji 133002, China.
Virol Sin. 2012 Jun;27(3):179-86. doi: 10.1007/s12250-012-3246-9. Epub 2012 Jun 9.
Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation. We developed a cost-effective, rapid, and highly sensitive one-step "triplex RT-PCR enzyme hybridization" assay for simultaneous detections of Japanese Encephallitis virus (JEV, Flaviviridae), Getah virus (GETV, Togaviridae), and Tahyna virus (TAHV, Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction. The analytical sensitivity of this assay was 1 PFU/mL for JEV, 10 PFU/mL for GETV, and 10 PFU/mL for TAHV. This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods. When "triplex RT-PCR enzyme hybridization" was applied to 29 cerebrospinal fluid (CSF) samples that were JEV-positive by normal RT-PCR assay, all samples were strongly positive for JEV, but negative for GETV and TAHV, demonstrating a good sensitivity, specificity, and performance at CSF specimen detection.
虫媒病毒对全球公共卫生和农业构成了严重威胁。快速、准确地鉴定虫媒病毒相关疾病的病毒因子对于流行病学监测和实验室研究至关重要。我们开发了一种具有成本效益、快速且高度敏感的一步式“三重 RT-PCR 酶杂交”检测方法，可同时检测日本脑炎病毒（JEV，黄病毒科）、基孔肯雅热病毒（GETV，披膜病毒科）和塔希纳病毒（TAHV，布尼亚病毒科），该方法使用三对引物在一个 RT-PCR 反应中扩增三个靶序列。该检测方法的分析灵敏度为 JEV 1PFU/mL、GETV 10PFU/mL 和 TAHV 10PFU/mL。与传统的血清学检测和单 RT-PCR 反应方法相比，该检测方法明显更快且更经济。当“三重 RT-PCR 酶杂交”应用于 29 份经常规 RT-PCR 检测呈 JEV 阳性的脑脊液（CSF）样本时，所有样本均强烈检测到 JEV，但未检测到 GETV 和 TAHV，表明该方法在 CSF 标本检测中具有良好的灵敏度、特异性和性能。
