89E-4 |
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B. GREENE1, J. Yu2, M. Ahmedna2, I. Goktepe2, and W. Willis3. (1) Human Environment and Family Sciences, North Carolina A&T State Univ., 161 Carver Hall, Greensboro, NC 27411, (2) Dept. of Human Environment & Family Sciences, North Carolina A&T State Univ., 161 Carver Hall, Greensboro, NC 27411, (3) Department of Human Environment and Family Science, North Carolina A&T State University, 161 Carver Hall, Greensboro, NC 27411 Campylobacteriosis caused by Campylobacter spp. has become one of the major causes of foodborne diseases worldwide. Campylobacter infections in human are mainly associated with the consumption of contaminated foods, especially poultry and water. Effective control and prevention of Campylobacteriosis is usually hindered by the lack of rapid and sensitive detection methods which are typically hampered by the small load of the bacteria in food products. The objectives of this study were to 1) develop a sensitive and rapid immunoassay for detection of Campylobacter in chicken carcasses, and 2) concentrate the bacteria to the detection limit of the optimized method. Campylobacter jejuni were cultured on Mueller-Hinton agar (containing 5% defibrinated sheep blood) at 37 °C under microaerophilic conditions. Brucella broth was used as sub-culture broth to give a viable bacterial count of 106-108 CFU/ml. Chicken wash was artificially contaminated with known numbers of C. jejuni (in the range 0-104 CFU/mL) then centrifuged at 4000-8000 g for 5-15 minutes. The pellet was re-suspended in Brucella enrichment broth, and incubated at 37 °C under microaerophilic condition for 1-3 hours before the immunodetection. A Sandwich-ELISA using Rabbit antibody to C. jejuni as capture antibody, HRP-labeled rabbit antibody to C. jejuni as detection antibody was optimized. The optimum concentrations for C. jejuni detection in chicken wash were capture and detection antibodies at 8-10 µg/ml and 1-2 µg/ml, respectively, and incubation at 37°C and 1 hour. Under these conditions, the optimized immunodetection protocol could detect C. jejuni at the concentration 104 CFU/ml or higher. Such levels were achieved by centrifugation and short time incubation for sample with low bacterial loads (<102 CFU/mL). The detection protocol required only 7 hours, including chicken carcasses washing, enrichment and immunoassay procedure. Therefore, optimized immunodetection developed in this study can be used for rapid monitoring of Campylobactor in poultry products.
Session 89E, Food Microbiology: Pathogens
2005 IFT Annual Meeting, July 15-20 - New Orleans, Louisiana |