Forsberg M, Tagle R, Madej A, Molina J R, Carlsson M A
Department of Clinical Chemistry, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala.
Acta Vet Scand. 1993;34(3):255-62. doi: 10.1186/BF03548189.
A radioimmunoassay for bovine (bLH), ovine (oLH) and porcine (pLH) luteinizing hormone was developed using a human 125ILH tracer from a commercial kit and a monoclonal antibody (518B7) specific for LH but with low species specificity. Standard curves demonstrated similar binding kinetics when bLH, oLH and pLH were incubated with tracer and antibody for 2 h at room temperature. A 30-min delay in the addition of the tracer gave sufficient sensitivity when analysing pLH. Separation of antibody-bound LH from free hormone was achieved by using second antibody-coated micro Sepharose beads. The assay was validated and the performance compared with that of an RIA currently in use for determination of bLH and oLH (coefficient of correlation: 0.99 and 0.98). Regardless of the standards used, intra-assay coefficients of variation were < 10% for LH concentrations exceeding 1 microgram/L. The inter-assay coefficients of variation were < 15%. The assay was used for clinical evaluation demonstrating the pre-ovulatory LH surge in two cyclic cows, LH pulsatility in an oophorectomized ewe and LH response to GnRH injection in a boar.
利用市售试剂盒中的人 125I 促黄体生成素（LH）示踪剂和对 LH 具有特异性但物种特异性较低的单克隆抗体（518B7），开发了一种用于检测牛（bLH）、羊（oLH）和猪（pLH）促黄体生成素的放射免疫分析方法。当 bLH、oLH 和 pLH 在室温下与示踪剂和抗体孵育 2 小时时，标准曲线显示出相似的结合动力学。在分析 pLH 时，示踪剂添加延迟 30 分钟可提供足够的灵敏度。通过使用包被二抗的微 Sepharose 珠实现抗体结合的 LH 与游离激素的分离。对该分析方法进行了验证，并将其性能与目前用于测定 bLH 和 oLH 的放射免疫分析方法进行了比较（相关系数分别为 0.99 和 0.98）。无论使用何种标准品，当 LH 浓度超过 1 微克/升时，批内变异系数均<10%。批间变异系数<15%。该分析方法用于临床评估，证实了两头周期性奶牛排卵前的 LH 峰、一只卵巢切除母羊的 LH 脉冲性以及一头公猪对 GnRH 注射的 LH 反应。
