POLYMERASE CHAIN-REACTION ASSAY COUPLED WITH FLUORESCENCE DETECTION ON MICROWELL PLATES FOR LISTERIA-MONOCYTOGENES IN FOODS

被引:14
作者
CANO, RJ
NORTON, DM
INZUNZA, AE
SANCHEZ, JG
OSTE, C
机构
[1] UNIV NACL AUTONOMA MEXICO,DEPT MICROBIOL & MED FAMILIAR,MEXICO CITY,DF,MEXICO
[2] MUNDITEST,E-07015 PALMA DE MALLORCA,SPAIN
[3] BECKMAN INSTRUMENTS INC,FULLERTON,CA 92634
关键词
LISTERIA MONOCYTOGENES; FLUORESCENCE DETECTION; PCR; DAIRY FOODS;
D O I
10.4315/0362-028X-58.6.614
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
This study evaluates a polymerase chain-reaction assay coupled with fluorescence detection (PCR-FD) in microwell plates for Listeria monocytogenes in dairy and food samples. Guanidinium thiocyanate/silica-extracted cultures and milk and dairy-food samples were used as templates for a PCR assay in microwell plates, with a primer pair that amplifies a 221 bp segment of the internal transcribed spacer (ITS), in the presence of 0.5 mu g/ml of ethidium bromide. Fluorescence of the PCR products was measured with a CytoFluor 2300 fluorescence measurement system (Millipore Corporation, Bedford, MA). The lowest level of detection of the assay was 10 to 100 CFU. A total of 326 food samples were tested, both by culture and by PCR-FD, The overall sensitivity of the PCR-FD assay was 95.2% and the specificity was 89.9%. Positive and negative predictive values were 74.8% and 99.4%, respectively. Based on the results obtained in this study it appears that the PCR-FD assay described here can be useful for screening a large number of milk and dairy-food samples for contamination by Listeria monocytogenes.
引用
收藏
页码:614 / 620
页数:7
相关论文
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