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

Research output: Contribution to journalArticle

389 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)133-141
Number of pages9
JournalDiseases of Aquatic Organisms
Volume25
Issue number1-2
Publication statusPublished - 1996
Externally publishedYes

    Fingerprint

Keywords

  • Detection
  • PCR
  • Penaeid shrimp baculovirus
  • PmNOBIII
  • White spot
  • WSBV

ASJC Scopus subject areas

  • Aquatic Science
  • Ecology

Cite this

Lo, C. F., Leu, J. H., Ho, C. H., Chen, C. H., Peng, S. E., Chen, Y. T., Chou, C. M., Yeh, P. Y., Huang, C. J., Chou, H. Y., Wang, C. H., & Kou, G. H. (1996). Detection of baculovirus associated with white spot syndrome (WSBV) in penaeid shrimps using polymerase chain reaction. Diseases of Aquatic Organisms, 25(1-2), 133-141.