Isolation, Detection and Genomic Differentiation of Vibrio cholerae and Vibrio parahaemolyticus in Bachok, Kelantan
نویسندگان
چکیده
The objective of this study was to determine the genetic diversity and virulence factors of Vibrio cholerae and V. parahaemolyticus from selected aquatic environments in Bachok, Kelantan, Malaysia. Out of 50 water samples, 33 were confirmed with the presence of presumptive V. cholerae and V. parahaemolyticus based on biochemical tests. Primers targeting toxR gene specific for V. cholerae (800bp) and V. parahaemolyticus (368bp), tdh (251bp) and trh (250bp) virulence genes were used in the PCR. Results of PCR showed that only 25 isolates of V. parahaemolyticus and 3 of V. cholerae had the toxR gene. None of the V. parahaemolyticus isolates had the trh and tdh genes. Repetitive extragenic palindromic PCR (REP-PCR) was used to determine the relatedness of these strains at the genomic level. Cluster analysis for REP-PCR showed 2 major clusters A-B, with several isolates from different locations grouped together indicating the geographical relatedness due to interconnected waterways. REPPCR showed that the species of Vibrios are genetically diverse. ABSTRAK Objektif kajian ini adalah untuk menentukan diversiti gen and faktor-faktor virulen Vibrio cholerae dan Vibrio parahaemolyticus yang diperolehi di sekitar perairan Bachok, Kelantan. Sejumlah 33 daripada 50 sampel air yang diuji melalui kaedah biokimia didapati positif untuk V. cholerae dan V. parahaemolyticus. Bagi amplifikasi PCR, primer toxR yang spesifik kepada V. cholerae (800bp) dan V. parahaemolyticus (368bp) serta primer-primer untuk mengesan gen-gen virulen seperti tdh (251bp) and trh (250bp) juga digunakan. Hasil PCR menunjukkan hanya 25 strain V. parahaemolyticus dan 3 strain V. cholerae mempunyai gen toxR. Tiada gen virulen dikesan pada mana-mana strain V. parahaemolyticus. REP-PCR diaplikasi untuk menentukan hubungan antara strain-strain yang telah diasingkan dalam kajian ini. Analisis dendrogram bagi REP-PCR menunjukkan 2 kumpulan utama, A-B. Strain-strain yang berasal dari lokasi yang berbeza dikumpulkan bersama. Ini mungkin disebabkan oleh faktor geografi di mana sistem perairan yang berkaitan. Keputusan REP-PCR menunjukkan variasi genetik yang nyata di dalam spesies Vibrio. (
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