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Simultaneous detection of three arboviruses using a triplex RT-PCR enzyme hybridization assay

  • Published:
Virologica Sinica

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 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.

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Authors and Affiliations

  1. Prevention of Veterinary Medicine Department, College of Agriculture, Yanbian University, Yanji, 133002, China

    Dan Dong & Tai-yuan Li

  2. State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China

    Dan Dong, Shi-hong Fu, Li-hua Wang, Zhi Lv & Guo-dong Liang

Authors
  1. Dan Dong
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  2. Shi-hong Fu
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  3. Li-hua Wang
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  4. Zhi Lv
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  5. Tai-yuan Li
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  6. Guo-dong Liang
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Corresponding author

Correspondence to Guo-dong Liang.

Additional information

Foundation items: NIH Grant (2U54AI057160-06); Development Grant of State Key Laboratory for Infectious Disease Prevention and Control (2008SKLID105).

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Dong, D., Fu, Sh., Wang, Lh. et al. Simultaneous detection of three arboviruses using a triplex RT-PCR enzyme hybridization assay. Virol. Sin. 27, 179–186 (2012). https://doi.org/10.1007/s12250-012-3246-9

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  • DOI: https://doi.org/10.1007/s12250-012-3246-9

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