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Distribution and Study on Single Nucleotide Polymorphism (SNP) of spvR Gene in Korean Isolates of Salmonella
Hyuk-Joon Kwon, D.V.M., Ph.D., Kyoung-Yoon Park, D.V.M., Ph.D., Joo-Youn Park, D.V.M. M.S. Yong-Ho Park, D.V.M., Ph.D., Sun-Joong Kim, D.V.M., Ph.D. and Han-Sang Yoo, D.V.M., Ph.D.
Department of Avian Diseases1, Department of Infectious Diseases3, Department of Microbiology4 College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul, Korea 2Clinical Pathology Laboratory, Bayer Korea Ltd
Vol.36 Num.6 (p335~340)
Background:The Salmonella virulence plasmid (spv) genes (spvR, A, B, C and D) on the large virulence plasmids of pathogenic Salmonella serotypes can replace the virulence of the whole plasmid. Recently, virulence plasmid-negative pathogenic Salmonella isolates were isolated. However, positive rates of spv genes among Korean Salmonella serotypes have been obscure. spv genes are conserved in compared to other virulence genes but there are single nucleotide polymorphisms (SNPs) conserved in only certain serotype. Such SNPs are useful for differentiation and understanding evolution of certain serotypes.
Materials and Methods:Salmonella serotypes isolated from live stocks [Salmonella typhimurium (ST, 26), S. enteritidis (SE, 10), S. gallinarum (SG, 40) and S. pullorum (SP, 53)] were used for colony-PCR. A primer set covering single nucleotide polymorphism (SNP) at 625th nucleotide of spvR was designed. The nucleotide sequences of amplicons were determined by cyclic sequencing method and RFLP was performed by using MseI.
Results:All isolates of SE, SG and SP, including four plasmid-negative isolates, showed specific amplicons but not all of ST (19/26, 73%) were positive to spvR. Based on the nucleotide sequence of 625th nucleotide and PCR-RFLP, SE, SG and SP [A(625)] and ST [G(625)] could be differentiated.
Conclusion:spvR can be used as a molecular marker to detect virulent SE, SG, SP and the SNP may be useful for differentiation of SE, SG, SP and ST. According to the SNP study SE may be evolutionarily closer to SG and SP than ST.
Keywords : Salmonella, spvR, Virulence plasmid, Single nucleotide polymorphism (SNP), Colony- PCR