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| 免疫磁捕获-实时荧光PCR快速检测鸡肉中沙门氏菌 | |
| 引用本文： | 张东方,袁飞,王娉,胡玥,陈颖,葛毅强.免疫磁捕获-实时荧光PCR快速检测鸡肉中沙门氏菌[J].食品与发酵工业,2011,37(8). |
| 作者姓名： | 张东方 袁飞 王娉 胡玥 陈颖 葛毅强 |
| 作者单位： | 1. 中国检验检疫科学研究院,北京100123／中国农业大学食品学院,北京100083 2. 中国检验检疫科学研究院,北京,100123 3. 中国农业大学食品学院,北京100083／中国农村技术开发中心,北京100045 |
| 基金项目： | 国家科技支撑计划项目(2009BAD9B06); 国家质检总局科研计划项目(2009IK312、2010IK178) |
| 摘 要： | 对免疫磁捕获-实时荧光PCR检测鸡肉中沙门氏菌进行了初步研究。主要包括免疫磁珠制备条件的选择及优化,通过实验最终确定了以6 mg/mL EDC和5 mg/mL NHS活化直径为700 nm的羧基聚苯乙烯磁性微球,与浓度为100μg/mL的沙门氏菌抗体在37℃振荡孵育1.5 h,制备得到抗沙门氏菌免疫磁珠,与市售沙门氏菌免疫磁珠试剂盒(Dynal产品)捕获效率相当。免疫磁捕获-实时荧光PCR检测方法检测限为104 CFU/mL左右,能在16 h内检测出鸡肉中104 CFU/mL的沙门氏菌,可用于食品中沙门氏菌的快速检测。 |
| 关 键 词： | 沙门氏菌 免疫磁捕获法 实时荧光PCR 检测 |
Rapid Detection of Salmonella in Chicken by Immunomagnetic Capture-Real Time PCR Assay |
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| Zhang Dong-fang,Yuan Fei,Wang Pin,Hu Yue,Chen Ying,Ge Yi-qiang.Rapid Detection of Salmonella in Chicken by Immunomagnetic Capture-Real Time PCR Assay[J].Food and Fermentation Industries,2011,37(8). | |
| Authors: | Zhang Dong-fang Yuan Fei Wang Pin Hu Yue Chen Ying Ge Yi-qiang |
| Affiliation: | Zhang Dong-fang1,2,Yuan Fei1,Wang Pin1,Hu Yue1,Chen Ying1,Ge Yi-qiang2,3 1(Institute of Food Safty,Chinese Academy of Inspection and Quarantine,Beijing 100123,China) 2(College of Food Science & Nutritional Engineering,China Agricultural University,Beijing 100083,China) 3(China Rural Technology Development Center,Beijing 100045,China) |
| Abstract: | In this preliminary study,the Immunomagnetic Beads Capture-Real time PCR(IMB-RTPCR) method for detecting Salmonella in food was established.The carboxyl magnetic microsphere with a particle diameter of 700 nm was selected as the solid phase carrier.EDC(6 mg/mL)and NHS(5 mg/mL)were used for activation.Subsequently,the 100 μg/mL anti-Salmonella antibody was incubated with the magnetic microsphere at 37℃ for 1.5 h in order to obtain the immunomagnetic bead.The capture efficiency of self-made beads was equivalent to Immunomagnetic beads kit(Dynal)while the former has lower cost.The method has the test limit up to 104 CFU/mL,and it could separate Salmonella effectively up to 104 CFU/mL within 16h.This method is applicable to routine food Salmonella testing. |
| Keywords: | Salmonella immunomagnetic capture method real time PCR detection |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |