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人绒毛膜促性腺激素的夹心型酶免疫测定法。

Sandwich-type enzyme immunoassay for human chorionic gonadotropin.

作者信息

Sekiya T, Furuhashi Y, Goto S, Kaseki S, Tomoda Y, Kato K

出版信息

J Endocrinol Invest. 1981 Jul-Sep;4(3):275-9. doi: 10.1007/BF03349444.

DOI:10.1007/BF03349444
PMID:7033340
Abstract

We have developed a sandwich-type enzyme immunoassay for human chorionic gonadotropin (hCG), in which antibody Fab'-beta-D-galactosidase complex and an antibody-immobilized silicone rubber solid phase were used. Despite the fact this assay system crossreacted about 40% with human luteinizing hormone (LH) that contains an immunologically very similar subunit to that of hCG, it proved to be highly sensitive with hCG measurable at levels as low as 0.3 mIU per assay tube. Using 25 microliter of urine sample, hCG levels in serum (10-1000 mlU/ml) or in urine (3-300 mlU/ml) could be determine with the same degree of precision as in radioimmunoassay without sample interference with the assay. The coefficients of variation in within-run, and between-run were 9.2-13.3%, and 4.2-18.8%, respectively. Values obtained with enzyme immunoassay correlate well with those of radioimmunoassay (r = 0.961, slope = 1.129, y-intercept = 3.7 mlU/ml for 35 serum samples) and hemagglutination assay (r = 0.954, slope = 0.951, y-intercept = 1.8 mlU/ml for 88 urine samples).

摘要

我们开发了一种用于人绒毛膜促性腺激素(hCG)的夹心型酶免疫测定法,其中使用了抗体Fab'-β-D-半乳糖苷酶复合物和固定有抗体的硅橡胶固相。尽管该测定系统与含有与hCG免疫上非常相似亚基的人促黄体生成素(LH)发生约40%的交叉反应,但事实证明它对hCG高度敏感,每个测定管中低至0.3 mIU的hCG水平都可检测到。使用25微升尿液样本,血清(10 - 1000 mIU/ml)或尿液(3 - 300 mIU/ml)中的hCG水平能够以与放射免疫测定相同的精密度进行测定,且样本不会干扰该测定。批内变异系数和批间变异系数分别为9.2 - 13.3%和4.2 - 18.8%。酶免疫测定获得的值与放射免疫测定(对于35个血清样本,r = 0.961,斜率 = 1.129,截距 = 3.7 mIU/ml)和血凝测定(对于88个尿液样本,r = 0.954,斜率 = 0.951,截距 = 1.8 mIU/ml)的值相关性良好。

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本文引用的文献

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THE SECOND INTERNATIONAL STANDARD FOR CHORIONIC GONADOTROPHIN.绒毛膜促性腺激素的第二项国际标准
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Immunoassay using antigen-enzyme conjugates.使用抗原-酶偶联物的免疫测定法。
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Radioimmunoassay: a method for human chorionic gonadotropin and human luteinizing hormone.放射免疫测定法:一种用于检测人绒毛膜促性腺激素和人促黄体生成素的方法。
Endocrinology. 1966 Jul;79(1):10-8. doi: 10.1210/endo-79-1-10.
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Studies on human chorionic gonadotrophin. I. Purification and some physico-chemical properties.人绒毛膜促性腺激素的研究。I. 纯化及某些物理化学性质
Acta Endocrinol (Copenh). 1968 Sep;59(1):89-104.
5
Radioimmunoassay for human chorionic gonadotropin.人绒毛膜促性腺激素放射免疫测定法
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A radioimmunoassay which specifically measures human chorionic gonadotropin in the presence of human luteinizing hormone.一种在人促黄体生成素存在的情况下特异性测量人绒毛膜促性腺激素的放射免疫分析方法。
Am J Obstet Gynecol. 1972 Jul 15;113(6):751-8. doi: 10.1016/0002-9378(72)90553-4.
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Immunoassay using antibody--enzyme conjugates.使用抗体-酶偶联物的免疫测定法。
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Competitive enzyme-liked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin.使用可溶性酶/抗体免疫复合物进行标记的竞争性酶联免疫测定法。I. 人绒毛膜促性腺激素的测定。
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Enzyme-linked immunoassay: conjugation of the Fab' fragment of rabbit IgG with beta-D-galactosidase from E. coli and its use for immunoassay.酶联免疫测定:兔免疫球蛋白G的Fab'片段与大肠杆菌β-D-半乳糖苷酶的偶联及其在免疫测定中的应用。
J Immunol. 1976 Jun;116(6):1554-60.
10
Use of rabbit antibody IgG-loaded silicone pieces for the sandwich enzymoimmunoassay of macromolecular antigens.使用负载兔抗体IgG的硅胶片进行大分子抗原的夹心酶免疫测定。
J Biochem. 1977 May;81(5):1557-66.