Development of a multiplex PCR system for the simultaneous detection of white spot syndrome virus and hepatopancreatic parvovirus infection. Issue 6 (16th October 2012)

• Record Type: Journal Article
• Authors: Jeeva, Subbiah; Kim, Nam‐Il; Jang, In‐Kwon; Choi, Tae‐Jin
• Is Part Of: Aquaculture research. Volume 45:Issue 6(2014:May)
• Page Start: 1073
• Page End: 1083
• Publication Date: 2012-10-16
• Subjects: Aquaculture -- Periodicals; Fishery management -- Periodicals; 639.8
• DOI: 10.1111/are.12045
• Languages: English
• ISSNs: 1355-557X

Abstract

<abstract abstract-type="main" id="are12045-abs-0001"> <title>Abstract</title> <p>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 <italic>Litopenaeus vannamei</italic>, <italic> Fenneropenaeus chinensis</italic> and <italic>Penaeus monodon</italic>. 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 <italic>L. vannamei</italic> 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 <italic>F. chinensis</italic> individuals showed 60% coinfection, and 40% were infected with HPV.</p> </abstract>