Date of Award:

5-2004

Document Type:

Thesis

Degree Name:

Master of Science (MS)

Department:

Nutrition, Dietetics, and Food Sciences

Department name when degree awarded

Nutrition and Food Sciences

Committee Chair(s)

Marie K. Walsh

Committee

Marie K. Walsh

Committee

Bart Weimer

Committee

Charlotte Brennand

Abstract

Salmonella is one of the leading foodborne pathogens causing illness today. Because of this, Salmonella rapid detection methods are under immense study for use in food. The traditional method, using the Food and Drug Administration- approved Bacterial Analytical Manual procedure, takes 4-6 days for Salmonella detection in food. Other rapid methods still take at least 16 h for detection due to their enrichment steps.

The hypothesis of this study was that the use of immobilized antibodies coupled with polymerase chain reaction (PCR) can be used for the rapid capture and detection of Salmonella spp. in food without the need for pre-enrichment. The rapid detection system was developed using immobilized anti-Salmonella antibody beads to capture and separate Salmonella from food without using an enrichment step. Detection of the immunocaptured Salmonella was done through PCR or an enzyme-linked immunosorbent assay (ELISA)-based system entitled Rapid Immuno-Capture (RIC).

The detection limit for buffer, chicken rinse, and shell eggs with static antibody capture PCR and RIC, was determined by inoculation of the Salmonella-free samples. The RIC assay detected Salmonella spp. in buffer at concentrations as low as 4 × 101 CFU/ml, and in chicken rinse and shell eggs, the assay detected Salmonella at 4 × 103 CFU/ml. The antibody capture with PCR detected Salmonella in buffer at concentrations as low as 4 × 102 CFU/ml, in chicken rinse at concentrations as low as 4 × 105 CFU/ml, and in shell egg at bacteria concentrations of 4 × 106 CFU/ml.

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