99A-29 |
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K. WAN and H. Wang. Dept. of Food Science & Technology, Ohio State Univ., 2015 Fyffe Ct., 110 Parker Food Science Bldg., Columbus, OH 43210-1007 Every year about 10% of our food supplies are lost due to spoilage. Spore-forming, thermophilic and acidophilic bacteria, molds and yeasts are the leading responsible microbial agents. Proper detection of the presence of these agents in raw materials and final products are critical for quality control. Conventional detection methods are time consuming and unsuitable for foods with limited shelf-life such as fruit juices. The objective of the study was to develop a rapid, specific and sensitive detection system for spoilage molds in juice products using a Taqman real-time PCR approach. The 18S rDNA regions were targeted for PCR primers-and-probe development. The 18S rDNA sequences from multiple fungi, selected fruits and meats found from the GenBank were aligned using the DNASTAR software package. Homology sequences were identified and conserved oligonucleotide primers for amplifying the large 18S rDNA region were derived. The 18S rDNA fragments from representative spoilage mold strains were amplified by PCR and cloned into the pCR2.1 vector. The nucleotide sequences of the cloned fragments were determined. A second alignment was performed including the newly identified sequences. The PCR primer-and-probe set fit the requirements for the Taqman system was developed. Spoilage molds were detected successfully from apple and orange juices by the Taqman PCR system using the developed primer-and-probe set. No cross-reaction was found with other common food-borne bacteria, yeasts and food ingredients. The whole detection procedures can be completed within a working day. This is a significant improvement compare to the current mainstream industry practices which take from 2 days to a couple of weeks from cultivation to organism characterization. The availability of this rapid mold detection system adds new value to the OSU CleanPlant package. Implementation of such a system can benefit the food industry by saving time and money, ensuring product quality and maintaining brand image.
Session 99A, Biotechnology: General
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