期刊
JOURNAL OF MOLECULAR DIAGNOSTICS
卷 12, 期 2, 页码 220-225出版社
ELSEVIER SCIENCE INC
DOI: 10.2353/jmoldx.2010.090036
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- Institut Colombiano pare el Desarrollo de la Ciencia y la Tecnologia, Francisco Jose de Caldas, Colciencias [3256-04-18105]
- Institut Nacional de Salud, Colombia
- Instituto Colombiano de Medicina Tropical-Universidad CES
A multiplex real-time polymerase chain reaction procedure was developed to identify the most prevalent clinical isolates of Salmonella enterica subsp. enterica. Genes from the rib, fliC, fljB, and viaB groups that encode the 0, H, and Vi antigens were used to design 15 primer pairs and TaqMan probes specific for the genes rfbJ, wzx, fliC, fljB, wcdB, the sdf-I sequence, and invA, which was used as an internal amplification control. The primers and probes were variously combined into six sets. The first round of reactions used two of these sets to detect Salmonella 0:4, 0:9, 0:7, 0:8, and 0:3,10 serogroups. Once the serogroups were identified, the results of a second round of reactions that used primers and probes for the flagellar antigen 1 genes, 1,2; e,h; g,m; d; e,n,x; and z(10), and the Vi gene were used to identify individual serovars. The procedure was standardized using 18 Salmonella reference strains and other enterobacteria. The procedure's reliability and sensitivity was evaluated using 267 randomly chosen serotyped Salmonella clinical isolates. The procedure had a sensitivity of 95.5% and was 100% specific. Thus, our technique is a quick, sensitive, reliable, and specific means of identifying S. enterica serovars and can be used in conjunction with traditional serotyping. Other primer and probe combinations could be used to increase the number of identifiable serovars. (J Mol Diagn 2010, 12:220-225: DOI: 10.2353/jmoldx.2010.090036)
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