Detection of baculovirus associated with white spot syndrome (WSBV) in penaeid shrimps using polymerase chain reaction

Chu Fang Lo, Jiann Horng Leu, Ching Hui Ho, Chau Huei Chen, Shao En Peng, You Tzung Chen, Chih Ming Chou, Pei Yan Yeh, Chang Jen Huang, Hsin Yiu Chou, Chung Hsiung Wang, Guang Hsiung Kou

研究成果: 雜誌貢獻文章同行評審

414 引文 斯高帕斯(Scopus)

摘要

White spot syndrome associated baculovirus (WSBV) is the causative agent of a disease which has recently caused high shrimp mortalities and severe damage to shrimp cultures. In this study, a strain of WSBV from black tiger shrimp Penaeus monodon was used to develop a diagnostic tool for the detection of WSBV and related agent infections in shrimp. The virions were purified from P. monodon infected with WSBV. Viral genomic DNA was extracted from purified virions by treating the virions with proteinase K and cetyltrimethylammonium bromide (CTAB) followed by phenol-chloroform extraction and ethanol precipitation. A qualitative assessment was performed using polymerase chain reaction (PCR) analysis on the viral DNA and primers specific to shrimp genomic DNA in order to monitor shrimp DNA contamination in the viral genomic DNA preparations. A WSBV genomic DNA library was constructed and, based upon the sequence of the cloned WSBV DNA fragment, we designed a WSBV-specific primer set for PCR to detect WSBV rnfection in penaeid shrimp. Samples which contained WSBV DNA yielded an evident amplification product showing the expected mobility of a 1447-bp DNA fragment, whereas nucleic acids extracted from tissue samples of clinically healthy shrimp showed no such DNA fragment, thereby confirming the specificity of our primers. By PCR with this primer set, it was demonstrated that the causative agents of white spot syndrome in different shrimp species are closely related. An effective diagnostic tool is thus provided for screening shrimp for WSBV infections, and may be important in preventing the further spread of this disease.
原文英語
頁(從 - 到)133-141
頁數9
期刊Diseases of Aquatic Organisms
25
發行號1-2
出版狀態已發佈 - 1996
對外發佈

ASJC Scopus subject areas

  • 海洋科學
  • 生態學

指紋

深入研究「Detection of baculovirus associated with white spot syndrome (WSBV) in penaeid shrimps using polymerase chain reaction」主題。共同形成了獨特的指紋。

引用此