Bioanalysis Research Group, Neurosciences Research Program, IMIM (Hospital del Mar Medical Research Institute), Barcelona, Spain;
Integrative Pharmacology and Systems Neuroscience Research Group, Neurosciences Research Program, IMIM (Hospital del Mar Medical Research Institute), Barcelona, Spain; Department of Pharmacology, Therapeutics and Toxicology and Department of Psychiatry, Universitat Autònoma de Barcelona (UAB), Cerdanyola del Vallès (Bellaterra), Spain; Clinical Pharmacology Unit. Hospital Universitari Germans Trias i Pujol-IGTP, Badalona, Spain;
Clin Chem. 2016 Oct;62(10):1353-60. doi: 10.1373/clinchem.2016.257592. Epub 2016 Aug 9.
Although it is being increasingly applied, blood collection for drug testing in sport presents some logistic issues that complicate full applicability on a large scale. The use of dried blood spots (DBS) could benefit compliant blood testing considerably owing to its simplicity, minimal invasiveness, analyte stability, and reduced costs. The aim of this study was to evaluate the applicability of DBS to the methodology approved by the World Anti-Doping Agency (WADA) for detection of doping by recombinant human growth hormone (rhGH) in serum.
A protocol for a single DBS analysis using the hGH isoforms differential immunoassays (kit 1 and kit 2) was developed and validated. A clinical study with healthy volunteers injected for 3 consecutive days with a low subcutaneous dose (0.027 mg · kg(-1) · day(-1) · person(-1)) of rhGH was conducted. Finger prick DBS and paired-time serum samples from arm venipuncture were compared.
The analysis of the DBS-based protocol indicated that with only a single blood spot it was possible to detect positivity for growth hormone abuse. In spite of the low rhGH dose administered and independently of the kit used, the window of detection for DBS was confirmed in all analyzed samples up to 8 h after rhGH administration and extended up to 12 h in 50% of the cases. Serum positivity was detected in all studied samples for 12 h after administration.
These results support the usefulness of DBS as a biological matrix for testing recent growth hormone abuse.
尽管血液采集用于运动中的药物检测的应用越来越广泛,但它仍存在一些后勤问题,这些问题使得大规模全面应用变得复杂。由于其简单性、微创性、分析物稳定性和降低的成本,使用干血斑(DBS)可以极大地受益于合规的血液检测。本研究的目的是评估 DBS 在世界反兴奋剂机构(WADA)批准的检测重组人生长激素(rhGH)在血清中兴奋剂的方法中的适用性。
开发并验证了一种使用 hGH 同工型差异免疫分析(试剂盒 1 和试剂盒 2)进行单次 DBS 分析的方案。对连续 3 天皮下注射低剂量(0.027mg·kg(-1)·day(-1)·person(-1))rhGH 的健康志愿者进行了临床研究。比较了手指刺破 DBS 和手臂静脉采血的配对时间血清样本。
基于 DBS 的分析方案表明,仅使用一个血斑就可以检测到生长激素滥用的阳性结果。尽管给予的 rhGH 剂量较低,并且与使用的试剂盒无关,在所有分析的样本中,DBS 的检测窗口在 rhGH 给药后 8 小时内得到确认,并且在 50%的情况下延长至 12 小时。在给药后 12 小时内,所有研究样本均检测到血清阳性。
这些结果支持 DBS 作为测试近期生长激素滥用的生物基质的有用性。