Application of randomly amplified polymorphic DNA markers (RAPD) to distinguish isolates of V.cholera and V.alginolyticus
V. Neela1, M.N. Shamsudin1*, W. Somarny1, R. Rozita2, A.R. Raha3 and S. Radu3
1Department of Clinical Laboratory Sciences and 2Department of Human Growth and Development, Faculty of
Medicine and Health Sciences; 3Department of Biotechnology, Faculty of Food Science and Biotechnology, University
Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
Received 10 August 2000 / Accepted 15 October 2000
Abstract.
A total of I I clinical isolates of V.cholera (9 Ogawa isolates, I Inaba isolate and I Bengal isolate) and 4 isolates of V.alginolyticus were studied by randomly amplified polymorphic DNA fingerprinting (RAPD) with twenty different 10 meroligonucleotide primers. Thirteen primers generated revealing a total of 185 DNA markers with sizes ranging from 100 10,000 bp which showed polymorphisms in all 15 isolates of V. cholera and V. alginolyticus. The overall percentages of the similarity ranged from 41 to 89% among V. cholera isolates and 21 to 43% for V. alginolyticus isolates. In addition, the genetic distance ranged from 0.320513 to 0.710843 for V.cholera and 0.275229 to 0.846774 for V. alginolyticus. The dendrogram showed that the V. alginolyticus isolates were grouped into one cluster and 2 single isolates, whereas V.cholera isolates were grouped into 2 clusters and 2 single isolates at a distance level of 70%. The three primers that produced distinct banding patterns for isolates of different Vibrio species were identified as being suitable primers to produce genetic markers, which have the potential to be used as species specific diagnostic probes. Our study therefore demonstrated that when suitable primers are identified and applied, RAPD analysis is simple and rapid method for clearly distinguishing between Vibrio spp.
Keywords: Vibrio cholera, vibrio alginolyticus, RAPD, dendrogram
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