应用ELISA和ELFA定量检测牛奶中葡萄球菌肠毒素A方法的建立  

Quantitative Detection of the Staphylococcal Enterotoxin A in Milk Using ELISA and ELFA Kit

作　　者：沈菊泉[1] 欧杰[2] 林露[1] 陈敏[3] 王婧[2] 胡洁云[1] 严维凌[1] SHEN Ju-quan1, OU Jie2 ,LIN Lu1, CHEN Min3, WANG Jing2 , HU Jie-yun1 , YAN Wei-ling1(1. Shanghai Food Res. Inst. , Shanghai 200235 ; 2. Coll. of Food Sci. ＆ Technol. , Shanghai Ocean Uni. , Shanghai 201306; 3. Shanghai Mun. Ctr. for Dis. Ctrl. ＆ Prevent. , Shanghai 20033)

机构地区：[1]上海市食品研究所,上海200235 [2]上海海洋大学食品学院,上海201306 [3]上海市疾病预防控制中心,上海200336

出　　处：《微生物学杂志》2018年第3期42-46,共5页Journal of Microbiology

基　　金：上海市科学技术委员会应用技术开发专项资金项目(2015-114)

摘　　要：探讨了Tecra SEs SET(ELISA法)和Vidas SET2(ELFA法)两种葡萄球菌肠毒素定性检测试剂盒用于定量检测牛奶中葡萄球菌肠毒素A(SEA)的可行性。根据GB/T27404-2008《实验室质量控制规范食品理化检测》的要求,对两种方法应用于定量检测时的检出限、校正曲线范围、相关系数和加标回收率指标进行了分析比较。实验结果显示,Tecra SEs SET对牛奶中SEA的检出限为0.79 ng/mL,校正曲线范围为0.79~10 ng/mL,相关系数r=0.997,SEA加标浓度为0.80、2.5和10 ng/mL时的回收率分别为110%、81%和100%。Vidas SET2的检出限为0.09 ng/mL,校正曲线范围为0.09~1.0 ng/mL,相关系数r=0.998,SEA加标浓度为0.1、0.25和1.0 ng/mL时的回收率分别为90%、95%和104%。上述结果结合对阳性样品的检测表明:这两种定性检测试剂盒能满足牛奶中SEA定量检测的要求。The aim of this study was to evaluate the feasibility for the quantitative detection of the staphylococcal enterotoxin A（ SEA） concentration in milk with two qualitative detection kits： Tecra SEs SET（ ELISA method） and Vidas SET2（ ELFA method）. The detection limit,range of calibration curve,correlation coefficient and spike recovery of both kits were compared and analyzed according to the requirements in GB/T27404-2008 ＂ Criterion on quality control of laboratories-Chemical testing of food＂. The experiment results showed that the detection limit of the Tecra Ses SET for the detection of SEA in milk was 0. 79 ng/mL with valid range of calibration curve ranged from 0. 79-10 ng/mL. The correlation coefficient was 0. 997. The spike recovery of Tecra Ses SET at the SEA concentration of 0. 80,2. 5 and 10 ng/mL were 110%,81%,and 100% respectively. The detection limit of Vidas SET2 was 0. 09 ng/mL with valid range of calibration curve ranged from 0. 09-1. 0 ng/mL. When correlation coefficient was 0. 998 the spike recoveries of Vidas SET2 at SEA concentration were 0. 1,0. 25,and 1. 0 ng/mL were 90%,95%,and 104% respectively. The above results combined with the detection of positive samples indicated that these two qualitative detection kits meet the requirement for quantitative detection of SEA in milk.

关 键 词：葡萄球菌肠毒素A 定量检测 酶联免疫 酶联荧光 

分 类 号：Q93-31[生物学—微生物学]