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Cai S.-H.,Guangdong Ocean University | Cai S.-H.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Cai S.-H.,Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institutes | Lu Y.-S.,Guangdong Ocean University | And 20 more authors.
Diseases of Aquatic Organisms | Year: 2013

The outer membrane proteins of Vibrio alginolyticus play an important role in the virulence of the bacterium and are potential candidates for vaccine development. In the present study, the ompW gene was cloned, expressed and purified. A DNA vaccine was constructed by inserting the ompW gene into a pcDNA plasmid. Crimson snapper Lutjanus erythropterus (Bloch) were injected intramuscularly with the recombinant plasmid pcDNA-ompW. The expression of the DNA vaccine was detected in gill, head kidney, heart, liver, spleen and injection site muscle of crimson snapper by RT-PCR 7 and 28 d post-vaccination. The ELISA results demonstrated that the DNA vaccine produced an observable antibody response in all sera of the vaccinated fish. In addition, crimson snapper immunized with the DNA vaccine showed a relative percentage survival (RPS) of 92.53%, indicating effective protection against V. alginolyticus infection. © 2013 Inter-Research.

Cao Y.T.,CAS South China Sea Institute of Oceanology | Cao Y.T.,Guangdong Ocean University | Cao Y.T.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Cao Y.T.,University of Chinese Academy of Sciences | And 6 more authors.
Letters in Applied Microbiology | Year: 2010

Aims: The purpose of this study was to establish a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive detection of Vibrio harveyi in mariculture shellfish. Methods and Results: A set of four primers, two outer and two inner primers, were designed from the toxR gene sequence of V. harveyi. The LAMP reaction was conducted at 65°C for 60 min. There were no cross-reactions with other bacterial strains indicating a high specificity of the LAMP. The detection sensitivity of the LAMP assay for V. harveyi with both of pure cultures and added shellfish cultures is about 10-5 dilution level (equivalent to 17·2 cells per reaction). The amplification products were detected by visual inspection using SYBR Green I. The detection sensitivity using the LAMP method was 10 times higher than that of conventional PCR. Conclusions: The LAMP assay established in this study is an extremely specific, sensitive and rapid for identification of V. harveyi in mariculture shellfish. Significance and Impact of the Study: This LAMP technique provides an important detecting tool for the detection of V. harveyi infection both in the laboratory and field. This technique is recommended as an applied protocol for health management programme and disease surveillance of in hatcheries as well as in grow-out pond, to prevent the disease outbreak. © 2010 The Society for Applied Microbiology.

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