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Development of a multiplex PCR system for the simultaneous detection of white spot syndrome virus and hepatopancreatic parvovirus infection

2012; Wiley; Volume: 45; Issue: 6 Linguagem: Inglês

10.1111/are.12045

1365-2109

Subbiah Jeeva, Nam-Il Kim, In‐Kwon Jang, Tae‐Jin Choi,

Insect symbiosis and bacterial influences

Aquaculture ResearchVolume 45, Issue 6 p. 1073-1083 Original Article Development of a multiplex PCR system for the simultaneous detection of white spot syndrome virus and hepatopancreatic parvovirus infection Subbiah Jeeva, Subbiah Jeeva Department of Microbiology, Pukyong National University, Busan, 608-737 Republic of KoreaSearch for more papers by this authorNam-Il Kim, Nam-Il Kim Molecular Science Division, Bioneer Corporation, Daejeon, 306-220 Republic of KoreaSearch for more papers by this authorIn-Kwon Jang, In-Kwon Jang National Fisheries Research & Development Institute, Incheon, 400420 Republic of KoreaSearch for more papers by this authorTae-Jin Choi, Corresponding Author Tae-Jin Choi Department of Microbiology, Pukyong National University, Busan, 608-737 Republic of KoreaCorrespondence: T-J Choi, Department of Microbiology, Pukyong National University, 45, Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea. E-mail: [email protected]Search for more papers by this author Subbiah Jeeva, Subbiah Jeeva Department of Microbiology, Pukyong National University, Busan, 608-737 Republic of KoreaSearch for more papers by this authorNam-Il Kim, Nam-Il Kim Molecular Science Division, Bioneer Corporation, Daejeon, 306-220 Republic of KoreaSearch for more papers by this authorIn-Kwon Jang, In-Kwon Jang National Fisheries Research & Development Institute, Incheon, 400420 Republic of KoreaSearch for more papers by this authorTae-Jin Choi, Corresponding Author Tae-Jin Choi Department of Microbiology, Pukyong National University, Busan, 608-737 Republic of KoreaCorrespondence: T-J Choi, Department of Microbiology, Pukyong National University, 45, Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea. E-mail: [email protected]Search for more papers by this author First published: 16 October 2012 https://doi.org/10.1111/are.12045Citations: 6Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract A multiplex PCR kit for simultaneous detection of white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) was developed and field testing was conducted. A 604-bp target sequence was selected from the vp28 gene of WSSV. A primer set was developed to amplify a 338-bp DNA fragment at the junction of the NS2 and NS1 protein genes of HPV after alignment of eight sequences from different strains. Another internal positive control primer set produced a 139-bp PCR fragment from the β-actin gene by alignment of this gene from Litopenaeus vannamei, Fenneropenaeus chinensis and Penaeus monodon. The detection limits, tested using purified plasmids, for WSSV and HPV were 21.4 and 19.0 copies respectively. The optimum ratio for HPV, WSSV and β-actin was 3:1:1, with an optimum annealing temperature of 57°C. Field test of the multiplex PCR with 170 L. vannamei individuals from 17 aquaculture farms showed 41.8% coinfection with WSSV and HPV, and 40.0% and 3.5% single infection with WSSV and HPV respectively. No virus-free shrimp farm was found. Ten wild catch F. chinensis individuals showed 60% coinfection, and 40% were infected with HPV. Citing Literature Volume45, Issue6May 2014Pages 1073-1083 RelatedInformation