DEVELOPMENT OF A REAL-TIME POLYMERASE CHAIN REACTION METHOD FOR THE DETECTION OF S. ENTERICA

Polymerase chain reaction (PCR) has become a widely used tool for the detection, identification and differentiation of pathogenic microorganisms in the diagnosis of human and animal diseases. Frequent outbreaks of Salmonella associated with food require the development of rapid detection methods to control the spread of the disease. Real-time PCR (RT-PCR) can detect the presence of pathogen DNA. Detection of Salmonella in RT-PCR was carried out using the developed primers SInv-1F, SInv-1R and the SE-Probe probe. Positive results were obtained when testing the specificity of the RT-PCR test system for the detection of S. enterica using DNA from the bacteria S. Enteritidis, S. Typhimurium and S. Virchow as templates. The developed RT-PCR test system for the detection of S. enterica DNA was tested on the DNA of heterologous microorganisms of the genera Pasterella, Clostridium, Escherichia coli, Bacillus, Staphylococcus, Pseudomonas, Klebsiella, Mycoplasma, Candida and Aspergillu. Heterologous microorganisms showed a negative result when testing the developed RT-PCR. The RT-PCR test system allows you to detect S. enterica DNA within 1-10 microbial cells. The developed RT-PCR test system made it possible to identify 99 (9.7%) S. enterica isolates as a result of examining 1020 biological samples (883 food samples and 137 clinical samples) collected in 2018-2019. The diagnostic efficiency of the S. enterica RT PCR test system was 100%.

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