TRS-based PCR as a potential tool for inter-serovar discrimination of Salmonella Enteritidis, S. Typhimurium, S. Infantis, S. Virchow, S. Hadar, S. Newport and S. Anatum

Abstract

Salmonella enterica subsp. enterica comprises a number of serovars, many of which pose an epidemiological threat to humans and are a worldwide cause of morbidity and mortality. Most reported food infection outbreaks involve the serovars Salmonella Enteritidis and Salmonella Typhimurium. Rapid identification to determine the primary sources of the bacterial contamination is important to the improvement of public health. In recent years, many DNA-based techniques have been applied to genotype Salmonella. Herein, we report the use of a manual TRS-PCR approach for the differentiation of the Salmonella enterica subspecies enterica serovars in a single-tube assay. One hundred seventy Salmonella strains were examined in this work. These consisted of serovars S. Enteritidis, S. Typhimurium, S. Infantis, S. Virchow, S. Hadar, S. Newport and S. Anatum. Five of the TRS-primers, N6(GTG)4, N6(CAC)4, N6(CGG)4, N6(CCG)4 and N6(CTG)4, perfectly distinguished the S. Enteritidis and S. Typhimurium serovars, and the N6(GTG)4 primer additionally grouped the other five frequently isolated serovars. In our opinion, the TRS-PCR methodology could be recommended for a quick and simple DNA-based test for inter-serovar discrimination of Salmonella strains.

Numerical interpretation of TRS-PCR profiling results for Escherichia coli strains isolated from patients with bacteriuria in Lodz region, Poland

“…In our laboratory, we have developed an effective method for the differentiation and genotyping of bacterial strains called TRS-PCR [15][16][17][18][19]. Trinucleotide repeat sequences are kind of microsatellites dispersed in genome of every organism.…”

“…Trinucleotide repeat sequences are kind of microsatellites dispersed in genome of every organism. This test was successfully used to detect associations between CGG-PCR and GTG-PCR fingerprints and specific E. coli properties such as phylogroups and virulence profiles [15][16][17][18][19]. The method is simple, fast, reproducible and inexpensive.…”

“…For each strain, three separate TRS-PCR profiles were generated based on the presence of CAC, GTG, and CGG motifs in used primers. Conditions for GTG-PCR and CGG-PCR reactions, primers sequences, electrophoresis, reproducibility analysis, determination of diversity indices and bioinformatic analyses were performed as described elsewhere [15][16][17][18][19]. Conditions for CAC-PCR were the same as for GTG-PCR (Table 1S).…”

“…The three separate fingerprint similarity analyses based on CAC-PCR, GTG-PCR and CGG-PCR profiles using Pearson correlation (optimization 1%, position tolerance 1%) and grouping according to the UPGMA algorithm were performed. The reproducibility analysis of CAC-PCR was performed as previously described for CGG and GTG [15][16][17][18][19] and obtained by comparing the three separate fingerprints from three different PCR reactions for the three chosen UPEC strains.…”

One-Step PCR Detection of Salmonella Pullorum/Gallinarum Using a Novel Target: The Flagellar Biosynthesis Gene flhB

“…Traditional Salmonella serotyping is conducted according to the White–Kauffmann–Le Minor scheme, which identifies the somatic (O) and flagellar (H) antigens based on the agglutination of bacteria with specific antisera (Majchrzak et al, 2014). Serotyping allows comparison with historical data, because it has been used for almost 70 years.…”