Application of a chromogenic medium and the PCR method for the rapid confirmation and characterization of vibro parahaemolyticus from seafoods

Chong, Wan Chin (2004) Application of a chromogenic medium and the PCR method for the rapid confirmation and characterization of vibro parahaemolyticus from seafoods. [Final Year Project Report] (Unpublished)

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Abstract

Thiosulfate-citrate-bile salts-sucrose (TCBS) Agar, a chromogenic medium and biochemical test were used to isolate, pre-confirm and identify the presence of 45 Vibrio parahaemolYlicus isolates from 15 samples of 3 different species offish (ikan kembung. ikan bawal and ikan sangeh) in the Kuching-Samarahan district between March 2003 and July 2003. Among the presumptive isolates, 18 isolates (40%) were randomly chosen to be analyzed using specific PCR amplification targeting the Ihermolabile (II) gene. All 18 isolates generated PCR products of 450 bp in size, thus indicating all presumptive isolates were indeed V. parahaemolYlicus. RAPD-PCR technique was later performed to fingerprint the isolates using primer GEN 15008. Bands ranging from 250 bp to 2500 bp were generated showing polymorphism among the isolates. Data from the PCR reaction were later used to construct a dendogram for c1ustal analysis. Finding shows that the 38 isolates belonged to 2 major clusters containing 7 and 31 isolates, respectively and the Simpson Index of Diversity calculated was 0.909.

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