促性腺激素的时间分辨荧光免疫测定(TR-FIA)

A Iwasawa, K Tomizawa, T Kato, K Wakabayashi, Y Kato
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引用次数: 8

摘要

时间分辨荧光免疫分析法(TR-FIA)是最近发展起来的一种非同位素免疫分析法,用于测定促性腺激素(LH和FSH)。用N1-(对异硫氰酸atobenzyl)-二乙烯三胺-N1, N2, N3, N3-四乙酸eu -螯合物(DTTA)标记羊LH或FSH,并使用事先包被第二抗体的96孔滴度板进行竞争结合测定,然后使用时间分辨荧光法进行测量。TR-FIA对标准羊LH (NIDDK, LH- i -3)或FSH (NIDDK, FSH- i -1)的灵敏度约为25 pg/50微升样品。该检测系统应用于猪促性腺激素的异源检测。用猪垂体前叶细胞原代培养培养基进行稀释试验,得到线性关系。31种不同猪样品LH和FSH测定的组内和组间变异系数较低,组内变异系数在3.5 ~ 8.1%之间,组间变异系数在1.7 ~ 13.1%之间。放射免疫分析法与TR-FIA的相关系数LH为0.945,FSH为0.978。用TR-FIA观察GnRH对LH和FSH释放的刺激。因此,这种非同位素TR-FIA提供了与传统ria一样好的灵敏度、重现性和准确性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Time-resolved fluoroimmunoassay (TR-FIA) of gonadotropins.

Time-resolved fluoroimmunoassay (TR-FIA), which has recently been developed as a non-isotopic immunoassay, was intended for assay of gonadotropins (LH and FSH). Ovine LH or FSH was labeled with N1-(p-isothiocyanatobenzyl)-diethylenetriamine-N1, N2, N3, N3-tetraacetic acid Eu-chelate (DTTA), and competitive binding assay was performed using a 96-weel titer plate previously coated with a second antibody, followed by measurement using time-resolved fluorometry. TR-FIA for standard ovine LH (NIDDK, LH-I-3) or FSH (NIDDK, FSH-I-1) had a sensitivity of about 25 pg/50 microliters sample. The assay system was applied to heterologous assay of porcine gonadotropins. Linearity was obtained by the dilution test using medium from primary culture of porcine anterior pituitary cells. Intra- and inter-assay coefficients of variation of LH and FSH determination in 31 different porcine samples were satisfactorily low, between 3.5 and 8.1% (intra-assay) and between 1.7 and 13.1% (inter-assay). Correlation coefficients between radioimmunoassay and TR-FIA were calculated to be 0.945 for LH and 0.978 for FSH. Stimulation of LH and FSH release with GnRH was observed by TR-FIA. This non-isotopic TR-FIA thus provides as good sensitivity, reproducibility and accuracy as conventional RIAs.

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