Watanabe H, Miyamoto A, Fukui Y
Laboratory of Animal Genetics and Reproduction, Obihiro University of Agriculture and Veterinary Medicine, Japan.
Reprod Fertil Dev. 1997;9(6):597-601. doi: 10.1071/r97037.
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.
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。
