title: Use of TaqMan® real-time PCR for rapid detection of Salmonella enterica serovar Typhi.
abstract: We evaluated the performances of a newly designed real-time polymerase chain reaction (PCR) assay using TaqMan® probes to detect Salmonella Typhi. TaqMan® real-time PCR assays were performed by designed primers and probe based on the staG gene for detecting S. Typhi. The specificity of the assay was evaluated on 15 Salmonella serovars. The analytical specificity was evaluated on 20 non-Salmonella microorganisms. The analytical sensitivity was assessed using decreasing DNA quantities of S. Typhi ATCC 19430. Finally the detection capability of the TaqMan® real-time PCR assay on isolates recovered from patients with Salmonella infections was compared to the conventional PCR assay. Only S. Typhi strain had positive results when subjected to the assay using Typhi-specific real-time PCR. No amplification products were observed in real-time PCR with any of the non-Salmonella microorganisms tested. The TaqMan® real-time PCR was more sensitive than the conventional PCR. In conclusion, we found that the easy-to-use real-time PCR assays were faster than conventional PCR systems. The staG-based TaqMan® real-time PCR assay showed to be specific and sensitive method for the safe and rapid detection of the S. Typhi.
authors.affiliation: Baqiyatallah University of Medical Sciences Molecular Biology Research Center Tehran Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Ranjbar
authors.affiliation: Baqiyatallah University of Medical Sciences Molecular Biology Research Center Tehran Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Naghoni
authors.affiliation: Shiraz University of Medical Sciences Prof. Alborzi Clinical Microbiology Research Center Shiraz Iran.
authors.forename: Shohreh
authors.initials: S
authors.lastname: Farshad
authors.affiliation: Baqiyatallah University of Medical Sciences Applied Virology Research Center Tehran Iran.
authors.forename: Hadi
authors.initials: H
authors.lastname: Lashini
authors.affiliation: Baqiyatallah University of Medical Sciences Molecular Biology Research Center Tehran Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Najafi
authors.affiliation: Ilam University of Medical Sciences Clinical Microbiology Research Center Ilam Iran.
authors.forename: Nourkhoda
authors.initials: N
authors.lastname: Sadeghifard
authors.affiliation: Department of Sciences for Health Promotion and Mother-Child Care 'G. D'Alessandro', University Palermo Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Mammina
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods* / Q000592:standards; D017931:DNA Primers / Q000737:chemistry; D005798:Genes, Bacterial; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012485:Salmonella typhi / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D014435:Typhoid Fever / Q000382:microbiology
chemical_list: D017931:DNA Primers
keywords: Salmonella Typhi; TaqMan® real-time PCR assay; polymerase chain reaction