作　　者： ; ; ; ; ; ;

机构地区： 广东海洋大学食品科技学院

出　　处： 《微生物学杂志》 2012年第6期7-11,共5页

摘　　要： 优化磁珠和副溶血性弧菌血清的偶联条件,并对其捕获性能进行评价。以考马斯亮蓝法测定磁珠偶联血清蛋白量,考察磁珠型号、粒径大小、活化剂的浓度、偶联温度、时间等对偶联效果的影响,并利用平板计数法评价免疫磁珠的捕获性能。选择S型磁珠,活化剂采用EDC和NHS溶液(浓度均为0.005 g/L)各0.1 mL,25℃、24 h进行偶联,血清蛋白和磁珠结合效率最高;在优化的条件下制备的免疫磁珠能特异性富集副溶血性弧菌,最低检测限为5 cfu/100 mL。利用优化的条件可以明显提高磁珠与血清蛋白的结合效率,制备的免疫磁珠具有较好的特异性,其检测限和准确度都大幅提高,为副溶血性弧菌免疫磁珠试剂盒的研制奠定了基础。 The coupling conditions of magnetic beads and Vibrio parahaemolyticus （ Vp ） serum protein were optimized, and its capturing performance was evaluated. Coomassie brilliant blue method was used to determine protein quantity of magnetic beads coupling serum, investigate the influence of magnetic beads model, particle diameter, and activator concentration, coupling temperature and time on the coupling effects, and evaluate the capturing performance of immunoma^netic beads with plate counting method. S-type magnetic beads was selected, and adopted EDC as activator and NHS solution （ both concentration were 0. 005 g/L） 0.1 mL each, and carried out coupling at 25 ℃ for 24 h, the syndesis effects of serum protein and magnetic beads was the highest; the immunomagnctic bead prepared under the optimized conditions could specifically enrich Vp, and the tested limit was at 5 cfu/100 mL. Therefore, using optimized conditions the syndesis efficiency of the magnetic beads and serum protein could be significantly promoted, and the immunomagnetic beads had a higher specificity and the testing limit as well as the accuracy was promoted by wide margin, thus laid a foundation for Vp immunomagnetic beads kit research and development.

关 键 词： 免疫磁珠 副溶血性弧菌 血清蛋白 偶联条件 富集

领　　域： [生物学]