Real-Time PCR Detection of Staphylococcus aureus in Milk and Meat Using New Primers Designed from the Heat Shock Protein Gene htrA Sequence

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 he...

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Bibliographic Details
Published in:Journal of food protection 2007-12, Vol.70 (12), p.2855-2859
Main Authors: CHIANG, Yu-Cheng, FAN, Chih-Ming, LIAO, Wan-Wen, LIN, Chien-Ku, TSEN, Hau-Yang
Format: Article
Language:English
Subjects:
Animals
Bacterial Proteins - genetics
Biological and medical sciences
Colony Count, Microbial
Consumer Product Safety
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Food Contamination - analysis
Food industries
Food Microbiology
Fundamental and applied biological sciences. Psychology
General aspects
Heat-Shock Proteins - genetics
Humans
Meat - microbiology
Methods of analysis, processing and quality control, regulation, standards
Milk - microbiology
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Species Specificity
Staphylococcus aureus - isolation & purification
Temperature
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Summary: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 10(3) 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.
ISSN:0362-028X
1944-9097
DOI:10.4315/0362-028x-70.12.2855