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A real-time PCR method for the detection of Salmonella enterica from food using a target sequence identified by comparative genomic analysis

Chen, Jing ; Zhang, Lida ; Paoli, George C. ; Shi, Chunlei ; Tu, Shu-I ; Shi, Xianming

International journal of food microbiology, 2010-02, Vol.137 (2), p.168-174 [Periódico revisado por pares]

Amsterdam: Elsevier B.V

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Título:
A real-time PCR method for the detection of Salmonella enterica from food using a target sequence identified by comparative genomic analysis
Autor: Chen, Jing ; Zhang, Lida ; Paoli, George C. ; Shi, Chunlei ; Tu, Shu-I ; Shi, Xianming
Assuntos: 5′-nuclease real-time PCR ; Animals ; Arachis hypogaea ; bacterial contamination ; Bacterial Proteins - genetics ; Bacteriological Techniques - methods ; Bioinformatics ; Biological and medical sciences ; Chickens ; DNA Primers - genetics ; food analysis ; food composition ; food contamination ; Food industries ; Food Microbiology ; food pathogens ; Fundamental and applied biological sciences. Psychology ; General aspects ; genomics ; Membrane Transport Proteins - genetics ; Methods of analysis, processing and quality control, regulation, standards ; microbial detection ; model food systems ; molecular sequence data ; nucleotide sequences ; Pathogen detection ; polymerase chain reaction ; Polymerase Chain Reaction - methods ; Polymerase Chain Reaction - standards ; real time polymerase chain reaction ; Reference Standards ; Salmonella ; Salmonella enterica ; Salmonella enterica - genetics ; Salmonella enterica - isolation & purification ; Salmonella enteritidis ; Salmonella typhimurium ; Sensitivity and Specificity ; sequence analysis ; Target mining
É parte de: International journal of food microbiology, 2010-02, Vol.137 (2), p.168-174
Notas: http://dx.doi.org/10.1016/j.ijfoodmicro.2009.12.004
http://hdl.handle.net/10113/40836
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Descrição: A 5′-nuclease real-time PCR assay using a minor groove binding probe was developed for the detection of Salmonella enterica from artificially contaminated foods. S. enterica-specific sequences were identified by a comparative genomic approach. Several species-specific target sequences were evaluated for specificity. A real-time PCR assay was developed targeting a nucleotide sequence within the putative type III secretion ATP synthase gene ( ssaN). An internal amplification control (IAC) probe was designed by randomly shuffling the target probe sequence and a single-stranded oligonucleotide was synthesized to serve as an IAC. The assay demonstrated 100% inclusivity for the 40 Salmonella strains tested and 100% exclusivity for 24 non- Salmonella strains. The detection limit of the real-time PCR assay was 41.2 fg/PCR with Salmonella Typhimurium genomic DNA and 18.6 fg/PCR using Salmonella Enteritidis genomic DNA; 8 and 4 genome equivalents, respectively. In the presence of a natural background flora derived from chicken meat enrichment cultures, the sample preparation and PCR method were capable of detecting as few as 130 Salmonella cfu/mL. Using the developed real-time PCR method to detect Salmonella in artificially contaminated chicken, liquid egg and peanut butter samples, as few as 1 cfu/10 g of sample was detectable after a brief (6 h) non-selective culture enrichment.
Editor: Amsterdam: Elsevier B.V
Idioma: Inglês

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A real-time PCR method for the detection of Salmonella enterica from food using a target sequence identified by comparative genomic analysis

Chen, Jing ; Zhang, Lida ; Paoli, George C. ; Shi, Chunlei ; Tu, Shu-I ; Shi, Xianming

Amsterdam: Elsevier B.V

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