氯霉素残留一步式化学发光酶免疫法的建立

A one-step mircorplate chemiluminescent enzyme immunoassay for detection of residual chloramphenicol

【目的】建立一种简便、快速、精确的氯霉素(CAP)残留检测方法。【方法】在常规两步式化学发光酶免疫法(CLEIA)的基础上,同时加入一抗和二抗,并进一步优化抗原包被条件、抗体工作浓度、竞争反应时间等试验参数,建立了检测CAP残留的一步式CLEIA。【结果】该方法最佳检测范围为0.045～128μg/L,检测灵敏度达3.16μg/L,最低检测限为0.01μg/L,可检出国际规定的CAP残留标准底限值以下的残留量。与常规两步法相比,在不影响检测限和灵敏度的前提下,可缩短反应时间1.5h。【结论】采用一步式CLEIA检测CAP残留,与常规两步式CLEIA的检测效果相近,且耗时短,更符合快速检测的要求。

[Objective]The present study was aimed to establish a convenient, rapid and accurate method for detection of residual chloramphenicol （CAP）. [Method]Based on conventional two-step chemiluminescent enzyme immunoassay （CLEIA）, the two antibodies were added to solution at the same time, and the antigen coating conditions, antibody concentration, competitive reaction timing, etc., were optimized to establish one-step competitive indirect chemiluminescent enzyme immunoassay. [Result]The established one-step CLEIA was utilized to detect the CAP, and the best detection was found in range of 0.045-128.000 μg/L with 3.16 μg/L detection sensitivity, and 0.01 μg/L lowest detection limit. This method could detect the CAP residual value even below the lowest limit of international standard. As compared to the two- step CLEIA, one-step CLEIA could lessen 1.5 hour of reaction time on the premise of no effects on detection limit and sensitivity. [Conclusion ]The established one-step CLEIA method produced the same quality of detection results for residual CAP as produced by two-step CLEIA, while it has advantage of taking lesser time compared to the later one.