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Diseases of Aquatic Organisms

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DAO 50:9-12 (2002)  -  doi:10.3354/dao050009

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

Jyh-Ming Tsai1,*, Lin-John Shiau1, Hsien-Hsiung Lee2, Peter W. Y. Chan1,2, Ching-Yu Lin2

1King Car Biotechnology Industrial Co., Ltd, Aquaculture Research and Development Center, No. 162-13 Qi Wu Lan Road, Jiao-Xi Xiang, I-Lan 266, Taiwan, ROC
2King Car Food Industrial Co., Ltd. Yuan-Shan Research Institute, No. 86 Chen-Hsiang Road, Yuan-San Xiang, I-Lan 264, Taiwan, ROC
*Present address: National Taiwan University, Department of Zoology, Taipei 106, Taiwan, ROC. E-mail:

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, 9195 F/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 internal control. 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 products were 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.

KEY WORDS: RT-PCR · WSSV · TSV · Penaeid shrimp

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