Use of TaqMan® real-time PCR for rapid detection of Salmonella enterica serovar Typhi

We evaluated the performances of a newly designed real-time polymerase chain reaction (PCR) assay using TaqMan® probes to detect Salmonella Typhi. TaqMan® real-time PCR assays were performed by designed primers and probe based on the staG gene for detecting S. Typhi. The specificity of the assay was evaluated on 15 Salmonella serovars. The analytical specificity was evaluated on 20 non-Salmonella microorganisms. The analytical sensitivity was assessed using decreasing DNA quantities of S. Typhi ATCC 19430. Finally the detection capability of the TaqMan® real-time PCR assay on isolates recovered from patients with Salmonella infections was compared to the conventional PCR assay. Only S. Typhi strain had positive results when subjected to the assay using Typhi-specific real-time PCR. No amplification products were observed in real-time PCR with any of the non-Salmonella microorganisms tested. The TaqMan® real-time PCR was more sensitive than the conventional PCR. In conclusion, we found that the easy-to-use real-time PCR assays were faster than conventional PCR systems. The staG-based TaqMan® real-time PCR assay showed to be specific and sensitive method for the safe and rapid detection of the S. Typhi

High prevalence of integron-mediated resistance in clinical isolates of Salmonella enterica

Salmonella enterica has become progressively resistant to antimicrobial agents worldwide as a result of genes carried on different classes of integrons. The aim of the current study was to investigate the molecular diversity of these integrons and their association with antimicrobial resistance in clinical S. enterica isolates from Tehran, Iran. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute. The presence of integrons was investigated by PCR using specific primers. Integrons were detected in 65 (47.1%) strains, with classes 1 and 2 being observed in 54 (39%) and 11 (8%) strains, respectively. Integron-positive isolates belonged to seven different S. enterica serovars, and all showed a multidrug-resistant (MDR) phenotype. Our findings show that integrons are widely disseminated among S. enterica strains from Tehran. Furthermore, the results that class 1 integrons were more prevalent than class 2 in Salmonella isolates, and that a statistical association with MDR patterns was observed, suggest that they are more likely to be important in conferring a resistant phenotype to Salmonella strains

Microbial diversity in the hypersaline Lake Meyghan, Iran

Lake Meyghan is one of the largest and commercially most important salt lakes in Iran. Despite its inland location and high altitude, Lake Meyghan has a thalassohaline salt composition suggesting a marine origin. Inputs of fresh water by rivers and rainfall formed various basins characterized by different salinities. We analyzed the microbial community composition of three basins by isolation and culturing of microorganisms and by analysis of the metagenome. The basins that were investigated comprised a green ~50 g kg−1 salinity brine, a red ~180 g kg−1 salinity brine and a white ~300 g kg−1 salinity brine. Using different growth media, 57 strains of Bacteria and 48 strains of Archaea were isolated. Two bacterial isolates represent potential novel species with less than 96% 16S rRNA gene sequence identity to known species. Abundant isolates were also well represented in the metagenome. Bacteria dominated the low salinity brine, with Alteromonadales (Gammaproteobacteria) as a particularly important taxon, whereas the high salinity brines were dominated by haloarchaea. Although the brines of Lake Meyghan differ in geochemical composition, their ecosystem function appears largely conserved amongst each other while being driven by different microbial communities