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Title: Multiplex quantitative PCR for detection of staphylococcus aureus in foods
Authors: Orawan Atthasopa
Nathinee Panvisavas
Soraya Chaturongakul
Keywords: Staphylococcus aureus;Multiplex quantitative RT-PCR;Nuc gene, mecA gene
Issue Date: 2014
Abstract: Based on Thai Department of Disease Control (DDC), Staphylococcus aureus has ranked third as a causative agent of food poisoning in Thailand. Staphylococcal food poisoning occurs when food contaminated with staphylococcal enterotoxin is ingested. The toxin accumulates as the bacterial contamination exceed approximately 105 colony forming unit per gram of food. The objective of this study is to detect S. aureus in food using rapid, sensitive, and specific multiplex quantitative PCR for detection of S. aureus. Hydrolysis probe and primer sets were designed to target conserved regions of specific genes which are nuc and mecA and the designed oligos were used in detection of different S. aureus strains. The probe of nuc and mecA genes were tested by qPCR with labeled Quasar 670 and TAMRA as reporter dye, respectively. Spiked milk was used as a model for detection of S. aureus in simple food matrix. The detection limit of nuc qPCR primer and probe set was at approximately 5,000 CFU/ml (equivalent to 15 pg of DNA).
Appears in Collections:Microbiology: International Proceedings
Plant Science: International Proceedings
MU: International Proceedings

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