Publication: A simple and rapid immunochromatographic test strip for detection of pathogenic isolates of Vibrio harveyi
0
0
Issued Date
2007
Resource Type
File Type
application/pdf
ISSN
1677012
Other identifier(s)
2-s2.0-36348932879
Rights Holder(s)
มหาวิทยาลัยศรีนครินทรวิโรฒ
Bibliographic Citation
Journal of Microbiological Methods. Vol 71, No.3 (2007), p.256-264
Suggested Citation
Sithigorngul P., Rukpratanporn S., Pecharaburanin N., Suksawat P., Longyant S., Chaivisuthangkura P., Sithigorngul W. A simple and rapid immunochromatographic test strip for detection of pathogenic isolates of Vibrio harveyi. Journal of Microbiological Methods. Vol 71, No.3 (2007), p.256-264. doi:10.1016/j.mimet.2007.09.012 Retrieved from: https://hdl.handle.net/20.500.14740/4272
Abstract
Mouse monoclonal antibodies (MAbs) and rabbit polyclonal antibody (PAb) against Vibrio harveyi were generated from immunization of mice and rabbits with highly virulent isolate of V. harveyi. Two MAbs specific to virulent isolates of V. harveyi were obtained and one of them (VH4) was selected to conjugate with colloidal gold as the detector antibody was laid on a sample pad. Rabbit polyclonal antibody was used as the capture antibody at the test line (T) and goat anti-mouse IgG antibody (GAM) was used as the capture antibody at the control line (C) of nitrocellulose strip. The ready-to-use strip was held in a plastic case and then stored in a desiccated plastic bag. A sample volume of 100 μl of bacterial suspension from various sources mixed with application buffer was applied to the sample chamber at one end of the strip and allowed to flow by chromatography through the nitrocellulose membrane to the other end. In test samples containing virulent isolates of V. harveyi, the bacteria would bind to the monoclonal antibody conjugated with colloidal gold and the resulting complex would be captured by the antibodies at the test line to give a reddish-purple band. Any unbound monoclonal antibody conjugated with colloidal gold moved across the test line would be captured by the GAM and form a band at the control line (C). In sample without V. harveyi or with V. harveyi below the limit (< 106 CFU/ml) of detection for the kit, only the control line band was observed. If the test sample was pre-enriched in tryptic soy broth (TSB) for 6 h before application to the strip, the sensitivity would increase to 1-10 CFU/ml which is comparable to that of PCR. This method could be used to detect pathogenic isolates of V. harveyi in pond water or infected shrimp in order to monitor and to reduce the risk of V. harveyi outbreak in the shrimp culture. The beneficial features of this kit are that simple, convenient and quick results (within 15 min) can be obtained without the requirement of sophisticated tools or special equipments and skills. © 2007 Elsevier B.V. All rights reserved.
Subject(s)
Polyclonal antibody
Pyroxylin
Antibody production
Article
Bacterium isolation
Chromatography
Nonhuman
Polymerase chain reaction
Priority journal
Shrimp
Test strip
Vibrio harveyi
Animals
Antibodies, Bacterial
Antibodies, Monoclonal
Chromatography
Gold Colloid
Immunoassay
Immunohistochemistry
Mice
Rabbits
Reagent Kits, Diagnostic
Reagent Strips
Vibrio
Bacteria (microorganisms)
Capra hircus
Decapoda (Crustacea)
Litopenaeus vannamei
Mus
Oryctolagus cuniculus
Vibrio harveyi
Pyroxylin
Antibody production
Article
Bacterium isolation
Chromatography
Nonhuman
Polymerase chain reaction
Priority journal
Shrimp
Test strip
Vibrio harveyi
Animals
Antibodies, Bacterial
Antibodies, Monoclonal
Chromatography
Gold Colloid
Immunoassay
Immunohistochemistry
Mice
Rabbits
Reagent Kits, Diagnostic
Reagent Strips
Vibrio
Bacteria (microorganisms)
Capra hircus
Decapoda (Crustacea)
Litopenaeus vannamei
Mus
Oryctolagus cuniculus
Vibrio harveyi