Simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV)by multiplex reverse transcription-polymerase chain reaction (RT-PCR) in Pacific white shrimp Penaeus vannamei

Diseases of Aquatic Organisms

Hsien-Hsiung Lee¹  Ching-Yu Lin¹  Peter W. Y. Chan¹  Lin-John Shiau¹  Jyh-Ming Tsai¹ 

关键词: RT-PCR;    WSSV;    TSV;    Penaeid shrimp;   

DOI  :  10.3354/dao050009

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: An assay using a single-tube, 1-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) was established for the simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV). Three primer sets, 9195F/9992 R, 94 F2/R2, and ITS F/28S R, were mixed at a ratio of 3:1:1 to amplify specific fragments of the TSV, WSSV, and Penaeus vannamei genome, respectively, in the RT-PCR reaction. Shrimp samples were experimentally infected with WSSV and TSV.PCR-amplified products detected in the nucleic acid extraction of shrimp pleopods produced 4 kinds of results. With no virus infection, 1 fragment of 892 base pairs (bp) was amplified from a ribosomal RNA gene by primer set ITS F/28S R as an internalcontrol. In samples only infected by WSSV or TSV, 2 fragments could be seen: either from WSSV (530 bp) plus the internal control or TSV (231 bp) plus the internal control, respectively. In cases of co-infection with both viruses, all 3 amplified productswere detected simultaneously. This study is the first report of Penaeus vannamei specimens co-infected with WSSV and TSV being detected using a PCR method via experimental infection.

【 授权许可】

Unknown