PCR法检测原料乳中金黄色葡萄球菌肠毒素A基因型

Detection of SEA in Raw Milk by PCR

通过建立PCR方法来检测原料乳中金黄色葡萄球菌肠毒素A基因型,优化其退火温度并验证了其敏感性、特异性。结果表明此方法在退火温度为58.7℃时扩增效果较为理想;灵敏度为1.357 pg/μL;以大肠杆菌、枯草芽孢杆菌、嗜热链球菌、志贺氏菌、沙门氏菌的基因组DNA为模板作为特异性检测对照组并分别进行PCR扩增反应,结果为阴性。同时对采集的乳样进行检测,乳房炎乳样中检测出了金葡菌肠毒素A基因,正常乳中没有检测出金葡菌,初步判断乳房炎是由金葡菌引起的。

This experiment was established to detect the SEA in raw milk by PCR. In this part, we optimize the annealing temperature and to verify its sensitivity and specificity. The results show that the annealing temperature is 58.7 ~C and the amplification effect is more ideal; the clear channel in expansion means sensitivity of this method is 1.357 pg/lxL; the genomic DNA of e. coli, bacillus subtilis, streptococcus thermophilus cultured, ambition, salmonella shut were used as a template for PCR amplification reaction respectively, and the results were negative. At the same time ,to test the acquisition samples,Staphylococcal enterotoxin A gene was detected in mastitis sample, and normal milk without Staphylococcus aureus. The result was preliminary judgement that mastitis induced by Staphylococcus aureus.