Development of a Diagnostic Prototype for the Detection of Salmonella by the LAMP Method and Investigation of Diagnostic Value of this Prototype

Author(s): Gulnur SERDAR, Oktay GENC

Salmonellosis is a contagious disease of humans and animals caused by Salmonella bacteria. The disease affects all animals. But young, pregnant, and nursing animals are more at risk. The main symptom is enteritis. But septicemia, abortion, arthritis, and respiratory diseases can also happen. The LAMP method stands out in the diagnosis of Salmonella. It is easy to use, gives quick results, and does not need an expert. LAMP relies on the isothermal strand-displacement activity of the Bsm (Bacillus subtilis) or Bst (Bacillus stereothermophilus) polymerase enzyme. The method is simple. The reaction components stay at the same temperature. All amplification and fixing occur in one step. In this study, the invasive A (invA) gene, which is widely used for identification of Salmonella at the genus level, was preferred. The LAMP reaction was done at 65ºC for 60 minutes. The results were seen by electrophoresis and with SYBR Green I dye. The LAMP test's detection limit was 3.66x102 cfu/ml for S. Enteritidis. The PCR and RT-PCR tests' limits were 3.66x104 cfu/ml and 3.66x103 cfu/ml. The specificity of the LAMP test was determined as 100%.