Molecular identification of Salmonella Typhimurium from village chickens based on invA and spvC genes

Research (Published online: 23-04-2020)

22. Molecular identification of Salmonella Typhimurium from village chickens based on invA and spvC genes

Mwanaisha Mkangara, Ernest R. Mbega and Musa Chacha

Veterinary World, 13(4): 764-767

ABSTRACT

Aim: This study aimed to identify Salmonella enterica serovars by polymerase chain reaction (PCR) based on virulence genes invasion A (inv A) and Salmonella plasmid virulence C (spvC).

Materials and Methods: DNA extraction of eight bacteria isolates was done using the PowerSoil® DNA Isolation Kit. The amplification of invA and spvC genes was done using conventional PCR. The positive PCR products were purified using the GeneJET Purification Kit and then sequenced using ABI 3730 XL automated genetic analyzer. The sequences obtained were compared for similarities with other Salmonella serovars deposited on the NCBI GenBank using BLASTN.

Results: Four out of eight samples were amplified by primers FS139/RS141 that target invA gene with products of about 284 bp, and three out of four of the same invA positive samples were also amplified by primers FSPV-1/RSPV-2 targeting spvC with a product of about 571 bp. One sample was not amplified by primers FSPV-1/RSPV-2 as it lacked virulence plasmid. Analysis of sequences indicated 100% homology with closely related serovars of S. enterica subspecies enterica serovar Typhimurium.

Conclusion: Salmonella Typhimurium that contained invA and spvC genes are pathogenic and virulent strains.

Keywords: invasive gene A, polymerase chain reaction, Salmonella plasmid virulence gene, Salmonella Typhimurium, sequencing.