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Title: FLUORESCENT REAL-TIME MOLECULAR (PCR) DETECTION OF ZOONOTIC BACTERIA AND VIRUSES IN BULK MILK.

Author
item Perdue, Michael
item Karns, Jeffrey
item Van Kessel, Jo Ann
item Higgins, James

Submitted to: International Meeting on Molecular Epidemiology and Evolutionary Genetics in Infectious Disease
Publication Type: Abstract Only
Publication Acceptance Date: 6/10/2002
Publication Date: 7/26/2002
Citation: Perdue, M.L., Karns, J.S., Van Kessel, J.S., Higgins, J.A. 2002. Fluorescent Real-Time Molecular (PCR) Detection of zoonotic bacteria and viruses in bulk milk. p.3. . International Meeting on Molecular Epidemiology and Evolutionary Genetics in Infectious Disease.

Interpretive Summary:

Technical Abstract: With the increased consumption of unpasteurized milk and milk products has come increased concern over the quality of bulk milk coming from the dairy farm. Further, the potential for purposeful addition of pathogens as a terrorist action at the farm or milk tanker level is a new consideration. To increase the capability of rapidly detecting zoonotic pathogens in bulk milk, we have evaluated fluorescent real-time PCR analysis of several pathogens in whole and skim milk. Using commercially available reagents, reagents developed at our laboratory, and a commercially available portable FRT-PCR machine (R.A.P.I.D., Idaho Technology), spiking experiments demonstrated that as few as 10 cfu/ml of Escherichia coli 0157:H7, 5 cfu/ml of Salmonella enterica, 10 spores/ml of Bacillus anthracis (Stern strain) and 1 pfu/ml of bovine enteroviruses were detectible by standard DNA and RNA extraction techniques from whole and skim milk. The complete extraction and PCR reaction can be carried out and results obtained in less than an hour, making it a feasible screening tool for bulk milk before processing. Preliminary screening of hundreds of bulk milk tank samples from the United States indicates however that the levels of any E. coli 0157:H7, Salmonella spp. and Listeria monocytogenes in bulk milk, while occasionally detectible after enrichment, are generally below the detection level of the FRT-PCR assay.