Real-Time PCR Detection of Staphylococcus aureus in Milk and Meat Using New Primers Designed from the Heat Shock Protein Gene htrA Sequence
Authors: Chiang, Yu-Cheng1; Fan, Chih-Ming1; Liao, Wan-Wen1; Lin, Chien-Ku1; Tsen, Hau-Yang2
Source: Journal of Food Protection®, Volume 70, Number 12, December 2007 , pp. 2855-2859(5)
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Abstract:
Staphylococcus aureus may cause foodborne disease outbreaks and staphylococcal infections and is one of the major causes of mastitis. Rapid and reliable methods for detection of this microorganism in milk and other foods are needed. In this study, we designed a primer set from the sequence of the heat shock protein gene htrA, a gene coding for high-temperature-requirement A (HtrA) protein, and used it for real-time PCR detection of S. aureus isolates: 16 reference strains and 40 strains isolated from food-poisoning cases. All strains tested generated positive results. Bacterial strains other than S. aureus, including strains of other Staphylococcus species, did not produce positive results. When this primer set was used for the real-time PCR detection of S. aureus in milk and meat samples without the preenrichment step, samples with target cell numbers greater than 103 CFU/ml or CFU/g could be detected, indicating the potential quantitative ability of this real-time PCR assay. With a 10-h preenrichment step, however, a detection limit of 1 CFU/ml or CFU/g could be obtained.Document Type: Research article
Affiliations: 1: Institute of Industrial Biotechnology, Hung Kuang University, 34 Chung-Chi Road, Shalu, Taichung County, 433, Taiwan, Republic of China 2: Department of Food Science and Nutrition, Hung Kuang University, 34 Chung-Chi Road, Shalu, Taichung County, 433, Taiwan, Republic of China
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