Costagliola S, Niccoli P, Florentino M, Carayon P
Laboratoire de Biochimie Endocrinienne et Métabolique, Unité 38 INSERM, Faculté de Médecine, Marseille, France.
J Endocrinol Invest. 1994 Jun;17(6):407-16. doi: 10.1007/BF03347727.
This report describes the results of the second part of the collaborative study organized by a working group sponsored by the Community Bureau of Reference of the European Community Commission. The whole study was designed to understand the causes of discrepancy among LH immunoassay methods. In the parent report we described the characteristics of LH monoclonal antibodies. In the present work we focused on the comparison of 11 commercially available monoclonal antibody based kits and one polyclonal antibody based RIA. Recovery experiments of the second IS 80/552 and of a related LH preparation showed that curve calibration differed among kits. The ratio between the LH recovery of the two most different kits was 2.30. LH concentrations were determined, using the 12 assay methods in the sera of prepubertal children (n = 46), normal women (n = 26) and men (n = 39) before and after LHRH stimulation, post-menopausal women (n = 29) and patients with renal failure (n = 71) or polycystic ovaries (n = 28). In children LH was detected in 0 to 80% of the sera depending on the kit. In adults, the mean LH concentration provided by the 12 kits varied from 6.0 to 13.55 IU/L showing a ratio of 2.26 between the two most different kits. A thorough statistical analysis allowed to distinguish two groups of kits. The first groups consisting of 6 kits provided results close to those obtained by the RIA. The 5 other kits misrecognized circulating LH in subjects with various clinical status. These 5 kits were the only ones to use, as labelled probes, monoclonal antibodies specific for the holohormone (anti-alpha beta) whereas the 6 other kits used, as labelled probes, monoclonal antibodies directed to the alpha (1 kit) or the beta-subunit (5 kits). In both groups, the coated monoclonal antibodies are directed to either the beta-subunit (1/6 and 2/5 kits) or the holohormone (5/6 and 3/5 kits). Taken together these data suggest that discrepancy among LH assay kits is related to variation in standard curve calibration and in epitope specificity of monoclonal antibodies used in the kits. These findings may prove to be instrumental to alleviate differences among LH assay methods.
本报告描述了由欧洲共同体委员会参考局赞助的一个工作组组织的合作研究第二部分的结果。整个研究旨在了解促黄体生成素（LH）免疫测定方法之间差异的原因。在之前的报告中，我们描述了LH单克隆抗体的特性。在当前工作中，我们重点比较了11种市售的基于单克隆抗体的试剂盒和1种基于多克隆抗体的放射免疫分析（RIA）方法。对第二种国际标准品80/552和一种相关LH制剂的回收实验表明，各试剂盒之间的曲线校准存在差异。两种差异最大的试剂盒的LH回收率之比为2.30。使用这12种测定方法对青春期前儿童（n = 46）、正常女性（n = 26）和男性（n = 39）在促性腺激素释放激素（LHRH）刺激前后、绝经后女性（n = 29）以及肾衰竭患者（n = 71）或多囊卵巢患者（n = 28）的血清中的LH浓度进行了测定。在儿童中，根据试剂盒的不同，0%至80%的血清中检测到了LH。在成年人中，这12种试剂盒提供的平均LH浓度在6.0至13.55 IU/L之间，两种差异最大的试剂盒之间的比值为2.26。全面的统计分析能够区分出两组试剂盒。第一组由6种试剂盒组成，其提供的结果与RIA方法获得的结果相近。另外5种试剂盒在各种临床状态的受试者中错误识别了循环中的LH。这5种试剂盒是仅有的将针对全激素（抗αβ）的特异性单克隆抗体用作标记探针的试剂盒，而另外6种试剂盒则将针对α亚基（1种试剂盒）或β亚基（5种试剂盒）的单克隆抗体用作标记探针。在两组中，包被的单克隆抗体均针对β亚基（1/6和2/5试剂盒）或全激素（5/6和3/5试剂盒）。综合这些数据表明，LH检测试剂盒之间的差异与标准曲线校准以及试剂盒中使用的单克隆抗体的表位特异性变化有关。这些发现可能有助于减少LH检测方法之间的差异。
