Production andcharacterization ofmonoclonal antibody againstPerkinsus olseniinundulated surf clams Paphia undulata
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Creator Sirirat Kaewsalabnil, Supannee Leethochavalit, Janjaras Watanachote, Kittinan Komolpis and Nanthika Khongchareonporn
Title Production andcharacterization ofmonoclonal antibody againstPerkinsus olseniinundulated surf clams Paphia undulata
Publisher Faculty of Agro-Industry
Publication Year 2558
Journal Title Food and Applied Bioscience
Journal Vol. 3
Journal No. 3
Page no. 231-238
Keyword Perkinsosis, Perkinsus olseni, Paphia undulata, monoclonal antibody, dot blotting
URL Website www.tci-thaijo.org/index.php/fabjournal
Website title www.tci-thaijo.org/index.php/fabjournal/article/view/78267
ISSN 2286-8615
Abstract Perkinsosis is a mollusk disease caused by protozoan parasite belonging to the genus Perkinsus. Perkinsosis has been found in some commercially important mollusks including oysters, clams and abalones. Heavy infection with Perkinsus often results in tissueinflammation and mass mortalities. To date, three stages have been described in the pathogen's life cycle: trophozoite, hypnospore and zoospore. The infections in mollusks cannot be diagnosed without specialized testing. Various methods have been applied in the diagnosis of these infections including the fluid thioglycollate medium (FTM) technique, immunological and PCR assays.In this study, monoclonal antibodies (MAb) for immunological assay against Perkinsus olseni were produced and characterized. Three ICR mice were immunized with P. olseni in the zoospore stage. The polyclonal antiserum showed the antibody titer of 1:128,000 by dot blotting. After three fusions of the splenocytes and myeloma cells, two hybridoma clones producing MAbs (2/B4/A2 and 8/H11/F2) against zoopores were obtained. The isotype of the MAbs 2/B4/A2 was IgG1 while that of 8/H11/F2 was IgM subclass. Only MAb 8/H11/F2 showed immunoreactivity with the zoospores and hypnospores stage. Both MAbs can be used to identify zoospores P. olseni from clams by dot blotting with the sensitivity range of 108109cell/ml.
Chiang Mai University

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