应用液相芯片检测空肠弯曲菌

摘要

Objective]The aim of the study is to establish liquid chip method for detection of Camplobacter jejuni. [Method] HIP protein monoclonal antibody againstCamplobacter jejuni was coupled with polystyrene microspheres and a liquid chip method was established for detection ofCamplobacter jejuni in combination with double antibody sandwich technique. The coupling rate of different antibody concentration with the microsphere was determined.The reaction conditions were optimized through L9(34)orthogonal design test,and the developed method was evaluated for its sensitivity and specificity. [Result] The results showed that the optimal test conditions were:the working poly-antibody concentration was 1:100,the working concentration of biotin labeled goat anti rabbit IgG was 1:500,the working concentration of SA-PE was10ug/mL and the reaction time of biotin labeled antibody and SA-PE was 90min. The sensitivity of the assay reached 103CFU/mL,without cross reaction with other common food borne pathogenic bacteria. [Conclusion]The method was of high sensitivity,specificity and reproducibility and could be used for rapid detection ofCamplobacter jejuni in food.%[目的]建立空肠弯曲菌液相芯片检测方法。[方法]本研究将空肠弯曲菌HIP蛋白单克隆抗体与聚苯乙烯微球偶联，结合双抗体夹心技术建立空肠弯曲菌液相芯片检测方法，测定不同浓度的抗体与微球偶联率。通过L9（34）正交设计试验优化方法的反应条件，并进行灵敏度和特异性试验。[结果]较优实验条件即多克隆抗体工作浓度为1：100、生物素标记的羊抗兔IgG工作浓度为1：500、SA-PE的工作浓度为10ug/mL、生物素标记的抗体与SA-PE反应时间为90min。该方法灵敏度可达103CFU/mL，与其他常见食源性致病菌无交叉反应。[结论]该方法灵敏度高、特异性强、重复性好，可快速检测食品中的空肠弯曲菌。