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目的建立一种可快速、精确地对蓖麻毒素(RT)进行定量检测的上转发光免疫检测技术(UPT-LF),即RT-UPT-LF。方法采用杂交瘤细胞技术制备蓖麻毒素单克隆抗体并对效价进行评价;采用效价最高的4种单抗分别与上转发光纳米颗粒(UCP-NP)共价偶联,分别作为检测带,进而两两配伍,确定最优的配伍组合条件建立RTUPT-LF方法;对RT-UPT-LF的敏感性、精密性、定量能力和特异性进行评价。结果与结论所建立的RT-UPT-LF方法可在15 min内完成对蓖麻毒素的检测,灵敏度可达0.5 ng/ml,定量范围为0.5~1000 ng/ml,与其他高浓度毒素无非特异反应,该法为蓖麻毒素检测提供了一种新手段。
OBJECTIVE: To establish a UPT-LF method for rapid and accurate quantitative detection of ricin (RT), RT-UPT-LF. Methods Monoclonal antibodies against ricin were prepared by hybridoma cell technology and their titer was evaluated. Four McAbs with the highest titer were covalently conjugated with UCP-NPs, respectively, and were used as detection bands , And then two to two compatibility, to determine the optimal combination of conditions to establish RTUPT-LF method; RT-UPT-LF sensitivity, precision, quantitative ability and specificity were evaluated. RESULTS AND CONCLUSION The RT-UPT-LF method was developed to detect ricin within 15 min with a sensitivity of 0.5 ng / ml and a quantitation range of 0.5-1000 ng / ml, which is non-specific to other high concentrations of toxins Reaction, the law provides a new means for the detection of ricin.