Cremonesi P, Perez G, Pisoni G, Moroni P, Morandi S, Luzzana M, Brasca M, Castiglioni B
Department of Biomedical Sciences and Technologies, University of Milan, Segrate, Milan, Italy.
Lett Appl Microbiol. 2007 Dec;45(6):586-91. doi: 10.1111/j.1472-765X.2007.02231.x. Epub 2007 Oct 4.
To develop an easy, rapid and efficient DNA extraction procedure for Staphylococcus aureus detection with a low number of steps and removing completely the PCR inhibitors, applicable to raw milk cheese samples, and to compare phenotypical and genotypical method to detect Staph. aureus isolates and staphylococcal enterotoxins (SEs) production.
A total of 33 bovine and caprine raw milk cheese samples were analysed by means of both classic microbiological and molecular techniques. All samples were positive for Staph. aureus contamination. The DNA extraction protocol optimized was found to achieve a detection limit of 100 CFU g(-1) for Staph. aureus. None of the samples tested with immunological assays contained SEs but in 14 of 33 samples a mixture of se positive (sea, sec, sed, seg, sel, sej) isolates were identified.
Staphylococcus aureus is a food-borne pathogen mainly detected in finished dairy products. The rapid and efficient detection of Staph. aureus isolates from dairy products is essential for consumer safety. The direct detection of pathogens from food is possible with careful attention to sample preparation and nucleic acid amplification optimization.
This study shows that raw milk cheese samples can be tested for Staph. aureus contamination with a rapid, simple and reproducible procedure.
开发一种简便、快速且高效的DNA提取方法,用于检测金黄色葡萄球菌,步骤少且能完全去除PCR抑制剂,适用于生乳奶酪样品,并比较检测金黄色葡萄球菌分离株和葡萄球菌肠毒素(SEs)产生的表型和基因型方法。
通过经典微生物学和分子技术对33份牛和羊的生乳奶酪样品进行了分析。所有样品的金黄色葡萄球菌污染均呈阳性。优化后的DNA提取方案对金黄色葡萄球菌的检测限为100 CFU g(-1)。免疫分析检测的样品中均未含有SEs,但在33份样品中的14份中鉴定出了se阳性(sea、sec、sed、seg、sel、sej)分离株的混合物。
金黄色葡萄球菌是一种主要在成品乳制品中检测到的食源性病原体。快速有效地检测乳制品中的金黄色葡萄球菌分离株对消费者安全至关重要。在仔细注意样品制备和核酸扩增优化的情况下,直接从食品中检测病原体是可能的。
本研究表明,生乳奶酪样品可以通过快速、简单且可重复的程序检测金黄色葡萄球菌污染。