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http://dx.doi.org/10.4014/jmb.1602.02050

A Modified Quantum Dot-Based Dot Blot Assay for Rapid Detection of Fish Pathogen Vibrio anguillarum  

Zhang, Yang (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
Xiao, Jingfan (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
Wang, Qiyao (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
Zhang, Yuanxing (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
Publication Information
Journal of Microbiology and Biotechnology / v.26, no.8, 2016 , pp. 1457-1463 More about this Journal
Abstract
Vibrio anguillarum, a devastating pathogen causing vibriosis among marine fish, is prevailing in worldwide fishery industries and accounts for grievous economic losses. Therefore, a rapid on-site detection and diagnostic technique for this pathogen is in urgent need. In this study, two mouse monoclonal antibodies (MAbs) against V. anguillarum, 6B3-C5 and 8G3-B5, were generated by using hybridoma technology and their isotypes were characterized. MAb 6B3-C5 was chosen as the detector antibody and conjugated with quantum dots. Based on MAb 6B3-C5 labeled with quantum dots, a modified dot blot assay was developed for the on-site determination of V. anguillarum. It was found that the method had no cross-reactivity with other than V. anguillarum bacteria. The detection limit (LOD) for V. anguillarum was 1 × 103 CFU/ml in cultured bacterial suspension samples, which was a 100-fold higher sensitivity than the reported colloidal gold immunochromatographic test strip. When V. anguillarum was mixed with turbot tissue homogenates, the LOD was 1 × 103 CFU/ml, suggesting that tissue homogenates did not influence the detection capabilities. Preenrichment with the tissue homogenates for 12 h could raise the LOD up to 1 × 102 CFU/ml, confirming the reliability of the method.
Keywords
Fish pathogen; quantum dot; rapid detection; Vibrio anguillarum;
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