Publication: Development of monoclonal antibodies specific to ToxA and ToxB of Vibrio parahaemolyticus that cause acute hepatopancreatic necrosis disease (AHPND)
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Issued Date
2017
Resource Type
File Type
application/pdf
ISSN
448486
Other identifier(s)
2-s2.0-85016138657
Rights Holder(s)
Scopus
Bibliographic Citation
Aquaculture. Vol 474, (2017), p.75-81
Suggested Citation
Wangman P., Chaivisuthangkura P., Sritunyalucksana K., Taengchaiyaphum S., Senapin S., Pengsuk C., Sithigorngul P., Longyant S. Development of monoclonal antibodies specific to ToxA and ToxB of Vibrio parahaemolyticus that cause acute hepatopancreatic necrosis disease (AHPND). Aquaculture. Vol 474, (2017), p.75-81. doi:10.1016/j.aquaculture.2017.03.039 Retrieved from: https://hdl.handle.net/20.500.14740/4171
Abstract
Toxin A (ToxA) and toxin B (ToxB) of Vibrio parahaemolyticus, which cause acute hepatopancreatic necrosis disease (AHPND), were prepared in a bacterial supernatant from Chinese isolates (CN-VPAHPND) by washing bacterial colonies off of TSA cultures. The supernatant was subsequently used in mouse immunization to produce monoclonal antibodies (MAbs). Three groups of MAbs were selected: one MAb specific to ToxA, two MAbs specific to ToxB and one MAb specific to V. parahaemolyticus (CN-VPAHPND). The MAbs specific to ToxA and ToxB recognized all 10 VPAHPND isolates from China, Vietnam, Malaysia and Thailand, but did not bind to the 20 non-VPAHPND isolates from various other sources, including Vibrio spp. and other bacteria. The MAbs specific to toxins were used to detect the recombinant proteins of His-tagged ToxA and GST-ToxB with sensitivities of 200fmolspot−1 and 10fmolspot−1, respectively, as determined by dot-ELISA. The MAbs were used to detect the toxins produced by bacteria or shrimp tissue lysate spiked with bacteria in a complex tissue sample at concentrations as low as 1CFUml−1 after pre-enrichment of the bacteria for 6h. The third group of MAb was specific to CN-VPAHPND isolate but did not recognize the other 9 out of 10 VPAHPND isolates from Vietnam, Malaysia and Thailand. However, this MAb demonstrated cross-reactivity to 1 out of 20 of the non-VPAHPND isolates and 3 out of 9 of the V. alginolyticus isolates. These findings indicate that the AHPND epidemic in Southeast Asia was not caused by the CN-VPAHPND isolate. The MAbs specific to ToxA and ToxB produced in this study could be used to detect both toxins directly by dot blotting. © 2017 Elsevier B.V.