title: Rapid molecular identification of Listeria species by use of real-time PCR and high-resolution melting analysis.
abstract: Identification of Listeria species via a molecular method is critical for food safety and clinical diagnosis. In this study, an assay integrating real-time quantitative PCR (Q-PCR) with high-resolution melting (HRM) curve analysis was developed and assessed for rapid identification of six Listeria species. The ssrA gene, which encodes a transfer-messenger RNA (tmRNA) is conserved and common to all bacterial phyla, contains a variable domain in Listeria spp. Therefore, Q-PCR and a HRM profile were applied to characterize this gene. Fifty-three Listeria species and 45 non-Listeria species were detected using one primer set, with an accuracy of 100% in reference to conventional methods. There was a 93.3% correction rate to 30 artificially contaminated samples. Thus, Q-PCR with melting profiling analysis proved able to identify Listeria species accurately. Consequently, this study demonstrates that the assay we developed is a functional tool for rapidly identifying six Listeria species, and has the potential for discriminating novel species food safety and epidemiological research.
authors.affiliation: Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, China. dazhi.jin@vanderbilt.edu
authors.forename: Dazhi
authors.initials: D
authors.lastname: Jin
authors.forename: Yun
authors.initials: Y
authors.lastname: Luo
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Weijia
authors.initials: W
authors.lastname: Fang
authors.forename: Julian
authors.initials: J
authors.lastname: Ye
authors.forename: Fang
authors.initials: F
authors.lastname: Wu
authors.forename: Gangqiang
authors.initials: G
authors.lastname: Ding
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D005516:Food Microbiology; D006801:Humans; D008087:Listeria / Q000235:genetics / Q000302:isolation & purification*; D008088:Listeriosis / Q000175:diagnosis; D012329:RNA, Bacterial / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D044366:Transition Temperature
chemical_list: D012329:RNA, Bacterial; C096222:tmRNA