Improved Template Preparation for PCR-Based Assays for Detection of Food-Borne Bacterial Pathogens

Applied and Environmental Microbiology - Tập 66 Số 10 - Trang 4539-4542 - 2000
Keith A. Lampel1, Palmer A. Orlandi1, Leroy Kornegay1
1Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, D.C. 20204

Tóm tắt

ABSTRACT Shigella flexneri , Salmonella enterica serotype Typhimurium, and Listeria monocytogenes were applied to FTA filters, and the filters were used directly as templates to demonstrate their sensitivity and applicability in PCR-based detection assays. With pure cultures, the sensitivities of detection by FTA filter-based PCR were 30 to 50 and 200 CFU for the gram-negative enterics and Listeria , respectively. Different numbers of S. flexneri cells were used in controlled contamination experiments with several different foods (produce, beef, and apple cider). Aliquots from concentrated food washes subsequently spotted onto FTA filters and assayed by PCR gave consistently positive results and detection limits similar to those observed with pure-culture dilutions. This universal method for PCR template preparation from bacterial cells is rapid and highly sensitive and reduces interference from food-associated inhibitors of PCR. In addition, its broad applicability eliminates the need for multiple methods for analysis of food matrices.

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Tài liệu tham khảo

10.1128/aem.57.12.3529-3534.1991

10.1016/0890-8508(90)90026-V

Belgrader P. Del Rio S. A. Turner K. A. Marino M. A. Weaver K. R. Williams P. E. Automated DNA purification and amplification from blood-stained cards using a robotic workstation. BioTechniques 19 1995 426 432

10.1006/mcpr.1998.0155

Burgoyne L. A. March 1996. Solid medium and method for DNA storage. U.S. patent 5 496 562.

10.1093/infdis/159.6.1126

10.1128/aem.59.5.1289-1293.1993

10.1111/j.1365-2672.1991.tb02951.x

10.1128/jb.174.13.4338-4349.1992

10.1128/jb.172.4.1905-1915.1990

10.4315/0362-028X-58.7.722

10.2144/00282pf01

10.3201/eid0505.990502

10.1128/JCM.38.6.2271-2277.2000

10.1006/abio.1997.2031

10.1016/S0043-1354(99)00071-8

10.1111/j.1472-765X.1994.tb00458.x

10.3201/eid0505.990503

10.1111/j.1472-765X.1990.tb00086.x

10.1046/j.1365-2672.1997.00300.x

10.1016/S0065-2423(08)60224-3

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Applied and Environmental Microbiology

Tập 66 Số 10

4539-4542

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