血清3,5,3’-三碘甲腺原氨酸化学发光定量免疫分析方法的建立

Quantitative Chemiluminescence Immunoassay for Serum Triiodothyronine

目的建立血清3,5,3’-三碘甲腺原氨酸（T3）化学发光定量免疫分析方法。方法采用固相竞争法,96孔微孔板包被T3抗体,辣根过氧化物酶标记T3,鲁米诺为发光底物,建立T3化学发光定量免疫分析方法。结果本方法的线性范围为0.5~8.0ng/m L;分析灵敏度为0.18ng/m L;批内变异系数为4.4%~6.6%,批间变异系数为5.5%~10.6%;回收率为90.0%~113.3%;高值T3血清样品经系列倍比稀释后的测定值与稀释度呈线性关系,相关系数为0.9922;与T4、γT3的交叉率分别为0.35%、0.54%;与罗氏电化学发光分析法的测定值具有较好的相关性。结论本方法具有分析灵敏度高、特异性好和操作简便快速的特点,适用于临床检测和科研应用。

Objective To establish a quantitative chemiluminescence immunoassay for triiodothyronine in human serum. Methods Competing method and the microplate coating technology was used in the assay. Anti-T3 antibody coated on the 96 microtiter plate, T3 labeled horse radish peroxidase, luminol and hydrogen peroxide were used as Luminescent substrate to establish the assay. Results The linear range of the assay was 0.5-8.0 ng/mL. The sensitivity of assay was 0. 18 ng/mL. The coefficients of variation for intra-assay were 4.4%- 6.6% ,and for inter-assay were 5.5%-10.6% respectively. The recovery rate of the samples was 90.0%- 113.3%. The correlation coefficient was 0. 9922 after serial dilutions of the high concentration sample. The cross-reactivity rate of thyronine and 3,3＇, 5＇-triiodothyronine （ yT3 ） with triiodothyronine was 0.35 % and 0.54% respectively. The examination result of the assay was correlated well with the Roche ECLIA. Conclusion The chemiluminescence immunoassay for triiodothyronine established in this study is sensitive, specific and convenient. It might be suitable for clinical and research application.