Title page for ETD etd-04142009-225600


Type of Document Dissertation
Author Datta, Shreya
URN etd-04142009-225600
Title Production and Application of Antibodies Produced against the Flagella and Hemolysin of Vibrio parahaemolyticus
Degree Doctor of Philosophy (Ph.D.)
Department Food Science
Advisory Committee
Advisor Name Title
Marlene Janes Committee Chair
Beilei Ge Committee Member
Paul Wilson Committee Member
Witoon Prinyawiwatkul Committee Member
Georgianna Tuuri Dean's Representative
Keywords
  • Vibrio parahaemolyticus
  • antibodies
Date of Defense 2009-04-03
Availability unrestricted
Abstract
Vibrio parahaemolyticus is a marine bacterium which is the leading cause of bacterial gastroenteritis due to consumption of raw or undercooked bivalve molluscan shellfish in United States and is considered an important seafood-borne pathogen throughout the world. The conventional methods for enumerating this bacteria from seafood is laborious and time consuming, hence the objective of our study was to develop antibody based method which would be sensitive, user-friendly and less time consuming to detect and enumerate Vibrio parahaemolyticus from oysters. In order to condense time and material two rapid and specific antibody based test methods were developed. In the first method, monoclonal antibodies were produced against the polar flagella of Vibrio parahaemolyticus. The monoclonal antibody was employed in Immunomagnetic separation (IMS) protocol and was then coupled with Real-time PCR (q-PCR). IMS coupled to q-PCR was able to detect bacterial numbers in less than 3 hrs with a sensitivity of 1,000 CFU/g of oyster homogenate. In the second method anti-hemolysin serum was produced and employed in Direct colony immunoblot (DCI) protocol to detect pathogenic Vibrio parahaemolyticus. DCI when compared with the currently used conventional method, DNA hybridization (DNAH) showed good correlation and was a faster and simpler method requiring less equipment and stringent laboratory conditions than DNAH. From our results we have found that IMS+q-PCR or DCI could be an useful addition to the methods available for enumerating pathogenic V. parahaemolyticus in seafood.
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