TY - JOUR
T1 - Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay
AU - Liu, Zongxiao
AU - Teng, Yong
AU - Liu, Hong
AU - Jiang, Yulin
AU - Xie, Xiayang
AU - Li, Huifang
AU - Lv, Jiangqiang
AU - Gao, Longying
AU - He, Junqiang
AU - Shi, Xiujie
AU - Tian, Feiyan
AU - Yang, Jingshun
AU - Xie, Congxin
N1 - Funding Information:
We are grateful to Dr. B. Hill (Centre for Environment, Fisheries & Aquaculture Science, UK) for providing IHNV, and Dr. R. Gudding (Norway National Veterinary Institute, Norway) for providing VHSV. This work was supported in part by China's General Administration for Quality Supervision, Inspection and Quarantine (AQSIQ) (2006IK003).
PY - 2008/4
Y1 - 2008/4
N2 - 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.
AB - 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.
KW - Fish rhabdoviruses
KW - Simultaneous detection
KW - mqRT-PCR
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U2 - 10.1016/j.jviromet.2007.12.017
DO - 10.1016/j.jviromet.2007.12.017
M3 - Article
C2 - 18299154
AN - SCOPUS:40849092317
SN - 0166-0934
VL - 149
SP - 103
EP - 109
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1
ER -