Simultaneous Detection of Three Arboviruses Using a Triplex RT-PCR Enzyme Hybridization Assay
10.1007/s12250-012-3246-9
- Author:
Dan DONG
;
Shihong FU
;
Lihua WANG
;
Zhi LV
;
Taiyuan LI
;
Guodong LIANG
- Publication Type:Journal Article
- Keywords:
Japanese Encephalitis virus(JEV);
Getah Virus(GETV);
Tahyna Virus(TAHV);
Multiplex RT-PCR;
Enzyme Hybridization
- From:
Virologica Sinica
2012;27(3):179-186
- CountryChina
- Language:Chinese
-
Abstract:
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 hybridizationassay 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,10PFU/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.
