一种利用生物素-链霉亲和素放大技术检测绵羊血浆中促卵泡激素的竞争性酶免疫测定法。

A competitive enzyme immunoassay for follicle-stimulating hormone in ovine plasma using biotin-streptavidin amplification.

作者信息

Watanabe H, Miyamoto A, Fukui Y

机构信息

Laboratory of Animal Genetics and Reproduction, Obihiro University of Agriculture and Veterinary Medicine, Japan.

出版信息

Reprod Fertil Dev. 1997;9(6):597-601. doi: 10.1071/r97037.

DOI:10.1071/r97037

PMID:9551663

Abstract

To determine follicle-stimulating hormone (FSH) concentration in ovine plasma, a second-antibody enzyme immunoassay using biotin-streptavidin amplification (BS-EIA) was developed and was validated by comparison with a routine radioimmunoassay (RIA). Ovine FSH was labelled with D-biotinyl-epsilon-aminocaproic acid-N-hydroxysuccinimide ester (biotin-7-NHS). The minimum and maximum detection limits were 0.40 ng mL(-1) and 25.0 ng mL(-1) respectively in the BS-EIA. Plasma samples from three superovulated ewes were measured for FSH by both BS-EIA and RIA. Correlation between values obtained by BS-EIA and RIA was 0.86 (n = 106), and the linear regression had a slope of 0.88 and an intercept of -0.07 (R2 = 0.74). The baseline of BS-EIA was slightly lower than that of RIA (0.69 ng mL(-1) v. 1.10 ng mL(-1)). The intra-assay and inter-assay coefficients of variation in BS-EIA were 13.8% and 16.7% (n = 106) respectively. The recovery rates were 104.8+/-2.7% (mean+/-s.e.m., n = 14). Changes in plasma FSH concentration from superovulated ewes evaluated by BS-EIA corresponded well with the results of routine RIA.

摘要

为了测定绵羊血浆中促卵泡激素（FSH）的浓度，我们开发了一种使用生物素-链霉亲和素放大技术的第二抗体酶免疫测定法（BS-EIA），并通过与常规放射免疫测定法（RIA）比较进行了验证。绵羊FSH用D-生物素基-ε-氨基己酸-N-羟基琥珀酰亚胺酯（生物素-7-NHS）进行标记。在BS-EIA中，最低检测限和最高检测限分别为0.40 ng mL⁻¹和25.0 ng mL⁻¹。用BS-EIA和RIA对三只超排母羊的血浆样本进行FSH测定。BS-EIA和RIA所得值之间的相关性为0.86（n = 106），线性回归的斜率为0.88，截距为-0.07（R² = 0.74）。BS-EIA的基线略低于RIA（0.69 ng mL⁻¹对1.10 ng mL⁻¹）。BS-EIA的批内和批间变异系数分别为13.8%和16.7%（n = 106）。回收率为104.8±2.7%（平均值±标准误，n = 14）。通过BS-EIA评估的超排母羊血浆FSH浓度变化与常规RIA的结果非常吻合。