甲状腺素运载蛋白时间分辨免疫荧光试剂盒的制备和效能评价

目的制备甲状腺素运载蛋白(transthyretin,TTR)的时间分辨免疫荧光(time-resolved fluoroimmunoassay,TRFIA)试剂盒并对其性能进行评价。方法将抗TTR的4H2单克隆抗体作为包被抗体包被至96孔板,用Eu^3+标记3E4单克隆抗体作为检测抗体,制备双抗体夹心TRFIA试剂盒,应用该试剂盒对42例川崎病(Kawasaki disease,KD)患儿及13名健康儿童的血清进行检测,以稀释回收率、准确度、精密度、稳定性等指标对试剂盒的效能进行评价。结果该研究制备的TTR TRFIA试剂盒与Western blot法同时检测42份KD临床样本和13份健康样本,两方法的结果符合率为100%,特异性强。试剂盒的最低检出浓度为0.05μg/ml,稀释回收率为88.00%~111.00%,分析内精密度为8.01%~9.17%,分析间精密度为8.88%~11.82%。稳定性实验表明该试剂盒可在4℃稳定保存6个月,37℃稳定保存7 d。结论该研究制备的TTR TRFIA试剂盒具有准确性高、方便快捷等优点,适用于大批量临床KD血清样品的TTR检测,为快速区分丙种球蛋白敏感型和无反应型的KD患儿提供了方法。

Use of fluorescent europium chelates as labels for detection of microcystin-LR in Taihu Lake, China

A method using the time-resolved fluorescence technology to establish a highly sensitive microcystin-LR （MC-LR） indirect com- petitive immunoassay was proposed in this work. This method was used to monitor the MC-LR level in source water and treated drinking water from Taihu Lake. Algae in the water samples were removed by eentrifugation, and the MC-LR level was quantified using this method. Testing results showed that the sensitivity of this method was 0.01 μg/L, and the dynamic measuring range was from 0.05 to 2 μg/L. The av- erage recovery was 115%, and the variation （CV） within and between different batches were 7.3% and 9.7%, respectively. Testing results also indicated that this time-resolved fluoroimmunoassay was sensitive and accurate in measuring MC-LR level, especially for quantitative analy- sis MC-LR level in bulk water.