AUTHOR=Xin Suhua , Zhu Hong , Tao Chenglin , Zhang Beibei , Yao Lan , Zhang Yaodong , Afayibo Dossêh Jean Apôtre , Li Tao , Tian Mingxing , Qi Jingjing , Ding Chan , Yu Shengqing , Wang Shaohui 

TITLE=Rapid Detection and Differentiating of the Predominant Salmonella Serovars in Chicken Farm by TaqMan Multiplex Real-Time PCR Assay

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=11

YEAR=2021

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2021.759965

DOI=10.3389/fcimb.2021.759965

ISSN=2235-2988

ABSTRACT=<p><italic>Salmonella</italic> has been known as an important zoonotic pathogen that can cause a variety of diseases in both animals and humans. Poultry are the main reservoir for the <italic>Salmonella</italic> serovars <italic>Salmonella</italic> Pullorum (<italic>S</italic>. Pullorum), <italic>Salmonella</italic> Gallinarum (<italic>S</italic>. Gallinarum), <italic>Salmonella</italic> Enteritidis (<italic>S</italic>. Enteritidis), and <italic>Salmonella</italic> Typhimurium (<italic>S</italic>. Typhimurium). The conventional serotyping methods for differentiating <italic>Salmonella</italic> serovars are complicated, time-consuming, laborious, and expensive; therefore, rapid and accurate molecular diagnostic methods are needed for effective detection and prevention of contamination. This study developed and evaluated a TaqMan multiplex real-time PCR assay for simultaneous detection and differentiation of the <italic>S</italic>. Pullorum, <italic>S</italic>. Gallinarum, <italic>S</italic>. Enteritidis, and <italic>S.</italic> Typhimurium. In results, the optimized multiplex real-time PCR assay was highly specific and reliable for all four target genes. The analytical sensitivity corresponded to three colony-forming units (CFUs) for these four <italic>Salmonella</italic> serovars, respectively. The detection limit for the multiplex real-time PCR assay in artificially contaminated samples was 500 CFU/g without enrichment, while 10 CFU/g after pre-enrichment. Moreover, the multiplex real-time PCR was applied to the poultry clinical samples, which achieved comparable results to the traditional bacteriological examination. Taken together, these results indicated that the optimized TaqMan multiplex real-time PCR assay will be a promising tool for clinical diagnostics and epidemiologic study of <italic>Salmonella</italic> in chicken farm and poultry products.</p>