AUTHOR=Xiong Dan , Song Li , Geng Shizhong , Tao Jing , An Shumin , Pan Zhiming , Jiao Xinan 

TITLE=One-Step PCR Detection of Salmonella Pullorum/Gallinarum Using a Novel Target: The Flagellar Biosynthesis Gene flhB

JOURNAL=Frontiers in Microbiology

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2016.01863

DOI=10.3389/fmicb.2016.01863

ISSN=1664-302X

ABSTRACT=<p><italic>Salmonella enterica</italic> serovar Pullorum/Gallinarum is an important infectious pathogen that has caused widespread problems for chicken industry. Traditional <italic>Salmonella</italic> serotyping is an expensive and time-consuming process. In this study, we developed a rapid one-step polymerase chain reaction (PCR) method to identify <italic>S.</italic> Pullorum/Gallinarum. The PCR-based assay focuses on <italic>flhB</italic>, which shows a deficient region only in <italic>S.</italic> Pullorum/Gallinarum, compared with that of other serovars. The specificity and sensitivity of the PCR system were evaluated. The developed PCR method could identify <italic>S.</italic> Pullorum/Gallinarum from 27 different <italic>Salmonella</italic> serovars and eight non-<italic>Salmonella</italic> pathogens. The minimum limit of DNA and the lowest number of cells of <italic>S.</italic> Pullorum for the PCR detection were no less than 5.85 pg/μL and 10 CFU, respectively. The method was applied to the analysis of <italic>Salmonella</italic> strains isolated from the chicken farm. The PCR-based testing results of the farm isolates were in concordance with those obtained using traditional serotyping method. This newly developed PCR-based system could be used to accurately screen for the presence of <italic>S.</italic> Pullorum/Gallinarum, and support traditional serotyping methods, especially in high-throughput screening situations.</p>