TITLE:
Development and Validation of SYBR Green-Based qPCR Technique of Detection and Quantification of Salmonella enterica
AUTHORS:
Rodica Sturza, Valentin Mitin, Irina Mitina, Alexandra Buga, Dan Zgardan, Emilia Behta
KEYWORDS:
Dilution, DNA, Primer, Real-Time PCR, Salmonella enterica
JOURNAL NAME:
Food and Nutrition Sciences,
Vol.12 No.11,
November
5,
2021
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.