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Research Article
Epidemiological Characterization of Salmonella gallinarum Isolates of Poultry Origin in India, Employing Two PCR Based Typing Methods of RAPD-PCR and PCR-RFLP

M. Habtamu Taddele, R. Rathore, K. Dhama and R.K. Agarwal

Asian Journal of Animal and Veterinary Advances, 2011, 6(11), 1037-1051.

Abstract

The present study was conducted to evaluate the discriminating power of RAPD-PCR and PCR-RFLP to characterize Salmonella gallinarum (S. gallinarum) isolates. Fifteen S. gallinarum isolates of poultry origin recovered from various sources in different regions of India including three reference strains were characterized using RAPD-PCR and PCR-RFLP. RAPD-PCR, using 5 primers (NSC I, NSC II, NSC III, URP-6 and primer 1290), grouped S. gallinarum isolates into 4-6 RAPD types. The discrimination power (D) of NSC II, URP-6 and primer 1290 was 0.848, 0.81 and 0.695, respectively and 0.62 for NSC I and NSC III. The phylogenetic dendrogram classified the isolates based on their geographic location. However, most S. gallinarum isolates were clustered closely together. In comparative analysis NSC II primer was placed in a separate cluster. The PCR amplification of the 16s rRNA and fimH genes produced approximately 572 bp and 1008 bp product, respectively, for all S. gallinarum isolates. Restriction Endonuclease (RE) enzymes viz., EcoRI, SmaI, AluI, BgiII, MspI and HaeIII and SalI, AluI, MspI and HaeIII were used to digest PCR products of 16s rRNA and fimH genes, respectively. These restriction enzymes generated 2-4 bands of varying molecular size ranging from 40 to 410 bp and 50 to 620 bp for 16s rRNA and fimH genes, respectively. AluI targeted against the fimH gene was the only enzyme which classified S. gallinarum isolates into three groups. In conclusion, the RAPD-PCR profile proved to be used as an epidemiological typing tool since it classified the S. gallinarum isolates according to their geographic origin.

ASCI-ID: 13-358

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