6533b824fe1ef96bd12814ea

RESEARCH PRODUCT

Use of TaqMan® real-time PCR for rapid detection of Salmonella enterica serovar Typhi

Caterina MamminaReza RanjbarAli NajafiShohreh FarshadAli NaghoniNourkhoda SadeghifardHadi Lashini

subject

SerotypeSalmonellaSettore MED/07 - Microbiologia E Microbiologia ClinicaBiologySalmonella typhimedicine.disease_causeReal-Time Polymerase Chain ReactionSettore MED/42 - Igiene Generale E ApplicataRapid detectionSensitivity and Specificitylaw.inventionlawTaqManmedicineHumansTyphoid FeverPolymerase chain reactionreal time typhoid fever diagnosisDNA PrimersGeneral Immunology and MicrobiologyGeneral MedicineSalmonella typhiVirologyMolecular biologyBacterial Typing TechniquesReal-time polymerase chain reactionSalmonella enterica serovar TyphiGenes Bacterial

description

We evaluated the performances of a newly designed real-time polymerase chain reaction (PCR) assay using TaqMan® probes to detectSalmonellaTyphi. TaqMan® real-time PCR assays were performed by designed primers and probe based on thestaGgene for detectingS.Typhi. The specificity of the assay was evaluated on 15Salmonellaserovars. The analytical specificity was evaluated on 20 non-Salmonellamicroorganisms. The analytical sensitivity was assessed using decreasing DNA quantities ofS.Typhi ATCC 19430. Finally the detection capability of the TaqMan® real-time PCR assay on isolates recovered from patients withSalmonellainfections was compared to the conventional PCR assay. OnlyS.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-Salmonellamicroorganisms 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. ThestaG-based TaqMan® real-time PCR assay showed to be specific and sensitive method for the safe and rapid detection of theS.Typhi.

10.1556/amicr.61.2014.2.3http://hdl.handle.net/10447/96701