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。

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.