Rapid molecular identification and differentiation of common Salmonella serovars isolated from poultry, domestic animals and foodstuff using multiplex PCR assay

Abstract

Salmonella is widely distributed throughout the world and can be found in poultry industry, animal breeding centers, food and feedstuffs of all geographical regions. This study was conducted to determine and identify Salmonella serovars isolated from poultry, calves and foodstuffs (poultry and animals products such as egg and meat). A total of one hundred isolates of Salmonella serovars including Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Gallinarum and Salmonella Pullorum consecutively were subjected to the conventional culture, biochemical and serological assays. The utility of molecular multiplex PCR was investigated to identify and differentiate among five Salmonella serovars which were identified according to the presence of rfbJ, fljB, invA, and fliC genes in S. Typhimurium, sefA, invA and spv genes in Salmonella Enteritidis, fljB, fliC and invA genes in Salmonella Infantis, hut and slgC genes in both Salmonella Gallinarum and Salmonella Pullorum and speC gene specifically in Salmonella Gallinarum. Biochemical assays and serotyping are complicated to directly differentiate between Salmonella Gallinarum and Salmonella Pullorum because of their antigenic similarity. According to the results, Multiplex PCR can be considered as simple, rapid, accurate and useful test to identify and differentiate among Salmonella serovars.

Characterization of Salmonella spp. isolated from chickens in Central China

“…After 16 h incubation at 37 °C, 0.1 mL and 10 mL of BPW were transferred into 10 mL of tetrathionate broth and 100 mL of selenite broth, respectively. Subcultures were spread on xylose-lysine-deoxycholate (XLD; Land Bridge, China) agar, xylose-lysine-tergitol 4 (XLT4; Land Bridge, China) agar, and Hektoen (HE; Hopebio, China) agar, and incubated at 37 °C for 24 h. The typical Salmonella colonies were confirmed by PCR amplification of the hut gene, and the primers were as follows, hut-F, 5′-ATGTTGTCCTGCCCCTGGTAAGAGA-3′, hut-R, 5′-ACTGGCGTTATCCCTTTCTCTGCTG-3′ [ 37 ]. Serotypes were determined by a slide agglutination test with O-antigen antiserum and a tube agglutination test with H-antigen antiserum, and confirmed by quadruplex PCR analysis as previously reported [ 38 ].…”

Quinolone resistance phenotype and genetic characterization of Salmonella enterica serovar Pullorum isolates in China, during 2011 to 2016

“…Swabs were cultured in 9 mL of Gram negative (GN) broth (Tianhe, China) at 37 °C for 24 h before aliquots of 100 mL of the broth were streaked onto Triple Sugar Iron agar (TSI, Oxoid, England). Typical Salmonella colonies were confirmed by PCR amplification of the hut gene, the primers were as followed, hut-F, 5’-ATGTTGTCCTGCCCCTGGTAAGAGA-3′, and hut-R, 5’-ACTGGCGTTATCCCTTTCTCTGCTG-3′ [18]. S .…”

DNA sequencing, genomes and genetic markers of microbes on fruits and vegetables

“…The genomes of approximately 20 strains of S. bongori have been registered, among which the representative strain is S. bongori NCTC 12419 (NCBI genome ID 1089). The sseL , spvC (Peterson et al , 2010), avrA , stn , stm (Amin et al , 2016), spv , hut , fljB (Alzwghaibi et al , 2018), hilA , fimA and hns (Jeyasekaran et al , 2011) gene regions have been reported as markers for the Salmonella genus. In addition, multiplex PCR analysis based on the fljB , gatD , invA , mdcA , stn and STM4057 genes is used for Salmonella genus identification (Lee et al , 2009).…”