Development and Validation of SYBR Green-Based qPCR Technique of Detection and Quantification of Salmonella enterica

Abstract

Salmonella enterica serovars is a leading cause of human gastroenteritis, and the incidence of salmonellosis is constantly increasing, causing millions of infections and many deaths annually. The detection of the pathogen in optimal terms is an essential factor for reducing the impact on the human body. In this work, SYBR Green I-based qPCR method of detection and quantification of Salmonella enterica was developed and validated. For detection of Salmonella enterica subsp. enterica, two pairs of primers were designed using publically available Primer-BLAST software. Primer efficiency was calculated by establishing a standard curve. The specificity, sensitivity, accuracy, and precision of PCR results were tested. Both primer pairs showed an acceptable performance, proving the developed techniques were sensitive, reliable and precise. The validated qPCR technology has a good potential to replace the traditional culture method in microbial diagnosis.