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

Abstract

ABSTRACT-White spot syndrome associated baculovlrus (WSBV) is the causative agent of a dlsease which has recently caused high shrimp mortahties and severe damage to shrimp cultures. In thls study, a strain of WSBV from black tiger shrimp Penaeus monodon was used to develop a diagnostlc tool for the detection of WSBV and related agent lnfect~ons in shnmp The vlnons were punfied from P monodon Infected with LVSBV V~ral genomlc DNA was extracted from purlfled vinons by treatlng the vlnons \ n t h proteinase K dnd cetyltnmethylammonium bromlde (CTAB) followed by phenol-chloroform extraction and ethanol precipitation A qualitative assessment Ivas performed using polymerase chain reaction (PCR) analys~s on the viral DNA and primers specif~c to shrimp genomic DNA in order to mon~t o r shrimp DNA contamination In the viral genomic DNA preparations A WSBV genomlc DNA llbrary was constructed and based upon the sequence of the cloned WSBV DNA fragment, we deslgned a LVSBV-specific prlmer set for PCR to detect WSBV Infection in penaeld shrimp Samples which contained WSBV DNA yielded a n evident ampl~f~catlon product showing the expected moblllty of a 1447-bp DNA fragment whereas n u c l e~c aclds extracted from tissue samples of clln~cally healthy shnmp showed no such DNA fragment, thereby confirming the speclficity of our pnmers By PCR with thls prlmer set, ~t was demonstrated that the causative agents of white spot syndrome in different shnmp specles are closely related An effective diagnostlc tool is thus provided for screening shnmp for \.VSBV infections, and may be important In preventing the further spread of this d~s e a s e KEY WORDS: WSBV . W h~t e s p o t . PmNOBIII . Detection . Penaeid shnmp baculovirus . PCR

Use of probiotic-supplemented diet on a Pacific white shrimp farm

“…The antimicrobial activity was estimated to be the reciprocal of the last well that inhibited bacterial growth. The presence of WSSV was determined by nested PCR, using the primers and conditions described by Lo et al (1996).…”

Molecular epidemiology of white spot syndrome virus within Vietnam

“…Since its discovery in China (Fujian) in 1991/1992, the virus spread quickly, causing serious losses to commercial shrimp farming worldwide (Cai et al, 1995;Flegel, 1997). The natural marine ecology is also threatened by WSSV as the virus has a wide host range, including salt and brackish water penaeids, crabs, spiny lobsters and freshwater shrimp and crayfish (Lo et al, 1996;Flegel, 1997;Wang et al, 1998;Chen et al, 2000;Maeda et al, 2000;Hameed et al, 2003). An overt clinical sign of infected shrimp is the presence of white spots on the exoskeleton (Chou et al, 1995).…”

Hsp70 expression in shrimp Litopenaeus vannamei in response to IHHNV and WSSV infection

“…Primers were designed from specific sequences of the viral genome, GenBank using the software OLIGO 3.0 as follow: initial denaturation step at 95°C for 5 min and then 35 cycles of 95°C for 30 s, 56°C for 30 s, and 72°C for 10 s. The set of primers used in the reaction were designed based in the sequence amplified by the nested PCR reaction WS146 F1 (5 0 -ACT-ACT-AAC-TTC-AGC-CTA-TCT-AG-3 0 ) and WS146 R1 (5 0 -TAA-TGC-GGG-TGT-AAT-GTT-CTT-ACG-3 0 ) internal prime pairs WS146 F2 (5 0 -GTA-ACT-GCC-CCT-TCC-ATC-TCC-A-3 0 ) and WS146 R2 (5 0 -TAC-GGC-AGC-TGC-TGC-ACC-TTG-T-3 0 ) [11,15]. The final reaction volume was 25 lL.…”