Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay

Zongxiao Liu, Yong Teng, Hong Liu, Yulin Jiang, Xiayang Xie, Huifang Li, Jiangqiang Lv, Longying Gao, Junqiang He, Xiujie Shi, Feiyan Tian, Jingshun Yang, Congxin Xie

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis.

Original languageEnglish (US)
Pages (from-to)103-109
Number of pages7
JournalJournal of Virological Methods
Volume149
Issue number1
DOIs
StatePublished - Apr 1 2008
Externally publishedYes

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Rhabdoviridae
Infectious hematopoietic necrosis virus
Novirhabdovirus
Carps
Real-Time Polymerase Chain Reaction
Viremia
Fishes
Viruses
Infectious pancreatic necrosis virus
Esocidae
Genes
Nucleoproteins
Glycoproteins
Cell Culture Techniques
Enzyme-Linked Immunosorbent Assay
RNA
Antigens
Sensitivity and Specificity
Research

Keywords

  • Fish rhabdoviruses
  • Simultaneous detection
  • mqRT-PCR

ASJC Scopus subject areas

  • Virology

Cite this

Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay. / Liu, Zongxiao; Teng, Yong; Liu, Hong; Jiang, Yulin; Xie, Xiayang; Li, Huifang; Lv, Jiangqiang; Gao, Longying; He, Junqiang; Shi, Xiujie; Tian, Feiyan; Yang, Jingshun; Xie, Congxin.

In: Journal of Virological Methods, Vol. 149, No. 1, 01.04.2008, p. 103-109.

Research output: Contribution to journalArticle

Liu, Z, Teng, Y, Liu, H, Jiang, Y, Xie, X, Li, H, Lv, J, Gao, L, He, J, Shi, X, Tian, F, Yang, J & Xie, C 2008, 'Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay', Journal of Virological Methods, vol. 149, no. 1, pp. 103-109. https://doi.org/10.1016/j.jviromet.2007.12.017
Liu, Zongxiao ; Teng, Yong ; Liu, Hong ; Jiang, Yulin ; Xie, Xiayang ; Li, Huifang ; Lv, Jiangqiang ; Gao, Longying ; He, Junqiang ; Shi, Xiujie ; Tian, Feiyan ; Yang, Jingshun ; Xie, Congxin. / Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay. In: Journal of Virological Methods. 2008 ; Vol. 149, No. 1. pp. 103-109.
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