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<p>Gene doping is known as the manipulation of congenital traits by gene therapeutic approaches with the intent of illicit athletic performance enhancement. A panel prototype suitable for multiplex gene doping detection by combining multiplex Polymerase Chain Reaction (PCR)-amplification with Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) analysis was developed and examined for its specificity and sensitivity, and its applicability in human sports drug testing programs was assessed. The panel comprises 20 assays for exon-exon-junction detection of seven human transgenes (<i>EPO, FST, GH1, IGF1, MSTN</i> (propeptide), <i>VEGFA, VEGFD</i>), which have been considered as material to routine doping controls, in one reaction. Alongside, a suitable reference material (RM) was designed and tested for its utility. An estimated LOD<sub>95</sub> of 1,500 cp / mL or 30 copies (cp) per reaction of the panel and 500 cp / mL or 10 cp per reaction of the RM was determined in plasmid-spiked human whole blood samples. The specificity and applicability of the panel and the RM was further determined by testing equine plasma samples obtained from an animal that received rAAV-delivered human transgenic <i>EPO</i> as well as 111 native human doping control samples.</p>

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Multiplex detection of seven transgenes for human gene doping analysis

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年6月20日

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  • Nana Naumann,
  • Carolin Do,
  • Caren Vollmert,
  • Maroje Krajina,
  • Andreas Thomas,
  • Hiu Wing Cheung,
  • Kin-Sing Wong,
  • Terence S. M. Wan,
  • Emmie N. M. Ho,
  • Mario Thevis

摘要

Gene doping is known as the manipulation of congenital traits by gene therapeutic approaches with the intent of illicit athletic performance enhancement. A panel prototype suitable for multiplex gene doping detection by combining multiplex Polymerase Chain Reaction (PCR)-amplification with Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) analysis was developed and examined for its specificity and sensitivity, and its applicability in human sports drug testing programs was assessed. The panel comprises 20 assays for exon-exon-junction detection of seven human transgenes (EPO, FST, GH1, IGF1, MSTN (propeptide), VEGFA, VEGFD), which have been considered as material to routine doping controls, in one reaction. Alongside, a suitable reference material (RM) was designed and tested for its utility. An estimated LOD95 of 1,500 cp / mL or 30 copies (cp) per reaction of the panel and 500 cp / mL or 10 cp per reaction of the RM was determined in plasmid-spiked human whole blood samples. The specificity and applicability of the panel and the RM was further determined by testing equine plasma samples obtained from an animal that received rAAV-delivered human transgenic EPO as well as 111 native human doping control samples.