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
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摘要
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. 0ng / m L. The sensitivity of assay was 0. 18 ng / m L. 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( γT3) with triiodothyronine was 0. 35% and0. 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.
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. 0ng / m L. The sensitivity of assay was 0. 18 ng / m L. 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( γT3) with triiodothyronine was 0. 35% and0. 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.
引文
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[8]官国英,刘中瑞,李丽波,等.白蛋白酶联免疫分析试剂盒的研制.同位素,2010,23(3):182-185.
[9]郭春祥,郭锡琼.介绍一种简单、快速、高效的辣根过氧化物酶标记抗体的过碘酸钠法.上海免疫学杂志,1983,3(2):97-100.
[10]李丽波,官国英,刘中瑞,等.β2-微球蛋白酶联免疫分析试剂盒的研制.同位素,2012,25(3):175-178.
[11]林珍,王栩,任世奇,等.化学发光酶免疫法测定游离三碘甲腺原氨酸.分析化学,2008,36(5):609-613.
[2]官国英,李丽波,刘中瑞,等.血清3,5,3’一三碘甲腺原氨酸酶联免疫分析试剂盒的研制.同位素,2011,29(2):98-101.
[3]潘中允.临床核医学.原子能出版社,1994:426-427.
[4]尹伯元,王仁芝,李振甲.标记免疫学.原子能出版社,1998:236-244.
[5]蔡锡麟,陈耀华,秦明秀.临床放射免疫学.原子能出版社,1994:133-138.
[6]吕建新,郑景晨.内分泌及代谢疾病的检验诊断.人民卫生出版社,2007:45-53.
[7]李龙,杜明华.检验核医学.东南大学出版社,2009:146-148.
[8]官国英,刘中瑞,李丽波,等.白蛋白酶联免疫分析试剂盒的研制.同位素,2010,23(3):182-185.
[9]郭春祥,郭锡琼.介绍一种简单、快速、高效的辣根过氧化物酶标记抗体的过碘酸钠法.上海免疫学杂志,1983,3(2):97-100.
[10]李丽波,官国英,刘中瑞,等.β2-微球蛋白酶联免疫分析试剂盒的研制.同位素,2012,25(3):175-178.
[11]林珍,王栩,任世奇,等.化学发光酶免疫法测定游离三碘甲腺原氨酸.分析化学,2008,36(5):609-613.