Primary Application and Establishment of a Duplex RT-PCR Method for CyHV2 and SVCV Detection

DOI

10.2991/meic-15.2015.262How to use a DOI?

Keywords

CyHV2; SVCV; RT-PCR;Electrophoresis

Abstract

To develop the rapid method for detecting of Spring viremia of carp virus (SVCV) and Cyprinid herpesvirus 2(CyHV2), 2 pairs of specific primers for duplex reverse transcriptase polymerase chain reaction (RT-PCR) were designed according to the nucleotide sequence information of CyHV2 and SVCV published in GenBank based on the construction of recombinant plasmids pMD19-T-SVCVG and pMD19-T-CyHV2, a duplex RT-PCR method were established via a serial of tests, including reaction temperature optimization test, sensitivity and specificity tests, and primary application tests. The results showed that the detection limits of the duplex RT-PCR for CyHV2 and SVCV detection were both approximately 80 copies of the cloned viral genomic fragments, as well as resulted in no cross-reaction for GCRV, Aeromonas veronii, Pseudomonas fluorescens, and Streptococcus isolated from fish, and the DNA(cDNA) of the normal Carassius auratus gibelio samples, and the application test results showed all of the 3 CyHV2 samples proved to be CyHV2 positive and there were not any amplification for the 15 clinical samples detection by the this method. The duplex RT-PCR established here is a rapid method with high specificity and sensitivity which are suitable for CyHV2 and SVCV detection.

Copyright

© 2015, the Authors. Published by Atlantis Press.

Open Access

This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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TY  - CONF
AU  - Yang Zexiao
AU  - Qiumei Xu
AU  - Yin Wang
AU  - Lirui Li
AU  - Xueping Yao
AU  - Kaiyu Wang
AU  - Ranyang Ren
AU  - Guili Li
AU  - Xingyu Lin
PY  - 2015/04
DA  - 2015/04
TI  - Primary Application and Establishment of a Duplex RT-PCR Method for CyHV2 and SVCV Detection
BT  - Proceedings of the 2015 International Conference on Mechatronics, Electronic, Industrial and Control Engineering
PB  - Atlantis Press
SP  - 1153
EP  - 1157
SN  - 2352-5401
UR  - https://doi.org/10.2991/meic-15.2015.262
DO  - 10.2991/meic-15.2015.262
ID  - Zexiao2015/04
ER  -