Abellan Rosario, Ventura Rosa, Palmi Ilaria, di Carlo Simonetta, Bacosi Antonella, Bellver Montse, Olive Ramon, Pascual Jose Antonio, Pacifici Roberta, Segura Jordi, Zuccaro Piergiorgio, Pichini Simona
Bioanalysis and Analytical Services Research Group, Neuropsychopharmacology Program, Institut Municipal d'Investigació Mèdica IMIM-Hospital del Mar, C/ Doctor Aiguader 88, 08003 Barcelona, Spain.
J Pharm Biomed Anal. 2008 Nov 4;48(3):844-52. doi: 10.1016/j.jpba.2008.05.037. Epub 2008 Jun 7.
Insulin-like growth factor-II (IGF-II), insulin-like growth factor binding proteins (IGFBPs) -2 and -3 and C-terminal telopeptide of type I collagen (ICTP) have been proposed, among others, as indirect biomarkers of the recombinant human growth hormone misuse in sport. An extended intra- and inter-laboratory validation of commercially available immunoassays for biomarkers detection was performed. ELISA assays for total IGF-II, IGFBP-2 and IGFBP-3 (IGF-II/ELISA1: DSLabs, IGFBP-2/ELISA2: Biosource, and IGFBP-3/ELISA3: BioSource) and an EIA assay for ICTP (ICTP/EIA: Orion Diagnostica) were evaluated. The inter- and intra-laboratory precision values were acceptable for all evaluated assays (maximum imprecision of 30% and 66% were found only for the lowest quality control samples of IGF-II and IGFBP-3). Correct accuracy was obtained for all inter-laboratory immunoassays and for IGFBP-2 intra-laboratory immunoassay. The range of concentrations found in serum samples under investigation was always covered by the calibration curves of the studied immunoassays. However, 11% and 15% of the samples felt below the estimated LOQ for IGF-II and ICTP, respectively, in the zone where lower precision was obtained. Although the majority of evaluated assays showed an overall reliability not always suitable for antidoping control analysis, relatively high concordances between laboratory results were obtained for all assays. Evaluated immunoassays were used to measure serum concentrations of IGF-II, IGFBP-2 and -3 and ICTP in elite athletes of various sport disciplines at different moments of the training season; in recreational athletes at baseline conditions and finally in sedentary individuals. Serum IGF-II was statistically higher both in recreational and elite athletes compared to sedentary individuals. Elite athletes showed lower IGFBP-2 and higher IGFBP-3 concentration with respect to recreational athletes and sedentary people. Among elite athletes, serum IGFBP-3 (synchronized swimming), and ICTP (rhythmic gymnastics) concentrations were sport-dependent. Over the training season, within athlete variability was observed for IGFBP-2 in case of taekwondo and IGFBP-2 and -3 in case of weightlifting. Variations due to those aspects should be taken in careful consideration in the hypothesis of setting reference concentration ranges for doping detection.
胰岛素样生长因子-II（IGF-II）、胰岛素样生长因子结合蛋白（IGFBP）-2和-3以及I型胶原C端肽（ICTP）等已被提议作为体育界重组人生长激素滥用的间接生物标志物。我们对用于生物标志物检测的市售免疫测定法进行了广泛的实验室内部和实验室间验证。评估了用于检测总IGF-II、IGFBP-2和IGFBP-3的ELISA测定法（IGF-II/ELISA1：DSLabs，IGFBP-2/ELISA2：Biosource，IGFBP-3/ELISA3：BioSource）以及用于检测ICTP的EIA测定法（ICTP/EIA：Orion Diagnostica）。所有评估测定法的实验室间和实验室内精密度值均可接受（仅在IGF-II和IGFBP-3质量最低的对照样品中发现最大不精密度分别为30%和66%）。所有实验室间免疫测定法以及IGFBP-2的实验室内部免疫测定法均获得了正确的准确性。所研究免疫测定法的校准曲线始终涵盖了所检测血清样品中的浓度范围。然而，在获得较低精密度的区域中，分别有11%和15%的IGF-II和ICTP样品浓度低于估计的定量下限。尽管大多数评估测定法总体上显示出可靠性，但并非总是适用于反兴奋剂控制分析，不过所有测定法在实验室结果之间都获得了相对较高的一致性。使用评估的免疫测定法测量了不同运动项目的精英运动员在训练赛季不同时间点、休闲运动员在基线条件下以及久坐不动个体血清中IGF-II、IGFBP-2和-3以及ICTP的浓度。与久坐不动个体相比，休闲运动员和精英运动员的血清IGF-II在统计学上均较高。与休闲运动员和久坐人群相比，精英运动员的IGFBP-2浓度较低，IGFBP-3浓度较高。在精英运动员中，血清IGFBP-3（花样游泳）和ICTP（艺术体操）浓度与运动项目有关。在训练赛季期间，跆拳道运动员的IGFBP-2以及举重运动员的IGFBP-2和-3在运动员个体之间存在差异。在设定兴奋剂检测参考浓度范围的假设中，应仔细考虑这些因素引起的变化。
