EVALUATION OF DIFFERENT DNA EXTRACTION PROCEDURES FOR THE DETECTION OF SALMONELLA FROM CHICKEN PRODUCTS BY POLYMERASE CHAIN-REACTION

被引:76
作者
SOUMET, C
ERMEL, G
FACH, P
COLIN, P
机构
[1] CNEVA,CENT RECH AVICOLE & PORCINE LAB,F-22440 PLOUFRAGAN,FRANCE
[2] LAB CENT HYG ALIMENTAIRE,PARIS,FRANCE
关键词
D O I
10.1111/j.1472-765X.1994.tb00458.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Polymerase chain reaction (PCR) was used after a short pre-enrichment culture to detect Salmonella subspecies in chicken fillets. A direct PCR assay performed with chicken meat inoculated with Salmonella Typhimurium produced no PCR products. Six different DNA extraction protocols were tested to recover efficiently Salmonella DNA after a short incubation period. Three of them gave results that were reliable, rapid and sensitive. Successful protocols used Proteinase K and/or a centrifugation step to concentrate the samples. For reliable detection of Salmonella subspecies, a few thousand bacterial cells per mi must be present. To obtain this number of bacterial cells with an inoculation of about one cell in 25 g of ionized food products, it was necessary to incubate samples for at least 10 h before PCR. A larger inoculum of approximately 10 cells in 25 g of ionized food products, required 8 h in culture broth to give positive results by PCR-based assay.
引用
收藏
页码:294 / 298
页数:5
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