引用本文: | 陈翠翠,梁焕坤,黎杰星,钟树海,李来庆,张立成.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 [J].中国临床新医学,0,():-. |
| CHEN Cui-cui,LIANG Huan-kun,LI Jie-xing,ZHONG Shu-hai,LI Lai-qing,ZHANG Li-cheng.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 [J].中国临床新医学,0,():-. |
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摘要: |
目的 制备甲状腺素运载蛋白(transthyretin,TTR)的时间分辨免疫荧光分析(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Eu3+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[确核表述
回答:修改无误,谢谢],应用该试剂盒对42例川崎病(KD)患儿及13名健康儿童的血清进行检测,以稀释回收率、灵敏度、精密度、稳定性等指标评价对试剂盒的性能进行评价。结果 本研究制备的TTR TRFIA试剂盒与Western Blot法同时检测42份川崎病临床样本和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0.05 μg/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%。稳定定性试验表明该试剂盒可在4 ℃稳定保存6个月,37 ℃稳定保存7天。结论 本研究制备的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。 |
关键词: 甲状腺素运载蛋白 川崎病 双抗体夹心 时间分辨免疫荧光分析 |
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基金项目:广东省科技计划项目(2016A010119067) |
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Development and effectiveness evaluation of time resolved immunofluorescence assay kit for detection of transthyretin |
CHEN Cui-cui,LIANG Huan-kun,LI Jie-xing,ZHONG Shu-hai,LI Lai-qing,ZHANG Li-cheng
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Guangzhou Youdi Biotechnology Co,Ltd,Guangzhou,China
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Abstract: |
Objective To prepare a kit of time-resolved fluoroimmunoassa(TRFIA) for detection of transthyretin (TTR), and evaluate its effectiveness. Methods The clone 4H2, the anti-TTR monoclonal antibody, was coated into 96-well microplate as the capture antibody, and the monoclonal antibody (clone 3E4) labeled with the Eu3+ as the detection antibody; which combined together to establish the TRFIA double-antibody sandwich method against TTR. Then, the kit was used to detect the TTR levels of 42 children with Kawasaki disease (KD) and 13 healthy children, and evaluated the efficiency by its dilution, recovery, sensitivity, precision and stability. Results Adopted the TTR TRFIA kit and Western blot method to detect 42 Kawasaki disease samples and 13 healthy samples at the same time, the coincidence rate between the two methods was 100%. The sensitivity of the prepared TRFIA kit is 0.05 μg/ml with the high specificity, and the dilution recovery rate is 88.00%~111.00%; also, the intra-assay precision of this method is 8.01%~9.17%, and the inter-assay precision is 8.88%~11.82%. In addition, it can be stably stored at 4 °C for half a year, and also at 37 °C for 7 days. Conclusion The TTR TRFIA kit developed has the advantages of high accuracy, convenience and quickness, which is suitable for the detection of large amount TTR clinical samples. Importantly, it provide a new method for differentiation of IVIG-sensitive and IVIG-non-responsive Kawasaki disease children. |
Key words: Kawasaki disease Transthyretin Double- antibody sandwich Time-resolved fluoroimmunoassa(TRFIA) |