Publication

A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA

de Boer, E. N., van der Wouden, P. E., Johansson, L. F., van Diemen, C. C. & Haisma, H. J., Aug-2019, In : Gene Therapy. 26, 7-8, p. 338-346 9 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

de Boer, E. N., van der Wouden, P. E., Johansson, L. F., van Diemen, C. C., & Haisma, H. J. (2019). A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA. Gene Therapy, 26(7-8), 338-346. https://doi.org/10.1038/s41434-019-0091-6

Author

de Boer, Eddy N ; van der Wouden, Petra E ; Johansson, Lennart F ; van Diemen, Cleo C ; Haisma, Hidde J. / A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA. In: Gene Therapy. 2019 ; Vol. 26, No. 7-8. pp. 338-346.

Harvard

de Boer, EN, van der Wouden, PE, Johansson, LF, van Diemen, CC & Haisma, HJ 2019, 'A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA', Gene Therapy, vol. 26, no. 7-8, pp. 338-346. https://doi.org/10.1038/s41434-019-0091-6

Standard

A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA. / de Boer, Eddy N; van der Wouden, Petra E; Johansson, Lennart F; van Diemen, Cleo C; Haisma, Hidde J.

In: Gene Therapy, Vol. 26, No. 7-8, 08.2019, p. 338-346.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

de Boer EN, van der Wouden PE, Johansson LF, van Diemen CC, Haisma HJ. A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA. Gene Therapy. 2019 Aug;26(7-8):338-346. https://doi.org/10.1038/s41434-019-0091-6


BibTeX

@article{9558ebddcc4347b2ae407b06bf3a5c1e,
title = "A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA",
abstract = "Gene doping confers health risks for athletes and is a threat to fair competition in sports. Therefore the anti-doping community has given attention on its detection. Previously published polymerase chain reaction-based methodologies for gene doping detection are targeting exon-exon junctions in the intron-less transgene. However, because these junctions are known, it would be relatively easy to evade detection by tampering with the copyDNA sequences. We have developed a targeted next-generation sequencing based assay for the detection of all exon-exon junctions of the potential doping genes, EPO, IGF1, IGF2, GH1, and GH2, which is resistant to tampering. Using this assay, all exon-exon junctions of copyDNA of doping genes could be detected with a sensitivity of 1296 copyDNA copies in 1000 ng of genomic DNA. In addition, promotor regions and plasmid-derived sequences are readily detectable in our sequence data. While we show the reliability of our method for a selection of genes, expanding the panel to detect other genes would be straightforward. As we were able to detect plasmid-derived sequences, we expect that genes with manipulated junctions, promotor regions, and plasmid or virus-derived sequences will also be readily detected.",
keywords = "gene doping, next generation sequencing",
author = "{de Boer}, {Eddy N} and {van der Wouden}, {Petra E} and Johansson, {Lennart F} and {van Diemen}, {Cleo C} and Haisma, {Hidde J}",
year = "2019",
month = "8",
doi = "10.1038/s41434-019-0091-6",
language = "English",
volume = "26",
pages = "338--346",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "7-8",

}

RIS

TY - JOUR

T1 - A next-generation sequencing method for gene doping detection that distinguishes low levels of plasmid DNA against a background of genomic DNA

AU - de Boer, Eddy N

AU - van der Wouden, Petra E

AU - Johansson, Lennart F

AU - van Diemen, Cleo C

AU - Haisma, Hidde J

PY - 2019/8

Y1 - 2019/8

N2 - Gene doping confers health risks for athletes and is a threat to fair competition in sports. Therefore the anti-doping community has given attention on its detection. Previously published polymerase chain reaction-based methodologies for gene doping detection are targeting exon-exon junctions in the intron-less transgene. However, because these junctions are known, it would be relatively easy to evade detection by tampering with the copyDNA sequences. We have developed a targeted next-generation sequencing based assay for the detection of all exon-exon junctions of the potential doping genes, EPO, IGF1, IGF2, GH1, and GH2, which is resistant to tampering. Using this assay, all exon-exon junctions of copyDNA of doping genes could be detected with a sensitivity of 1296 copyDNA copies in 1000 ng of genomic DNA. In addition, promotor regions and plasmid-derived sequences are readily detectable in our sequence data. While we show the reliability of our method for a selection of genes, expanding the panel to detect other genes would be straightforward. As we were able to detect plasmid-derived sequences, we expect that genes with manipulated junctions, promotor regions, and plasmid or virus-derived sequences will also be readily detected.

AB - Gene doping confers health risks for athletes and is a threat to fair competition in sports. Therefore the anti-doping community has given attention on its detection. Previously published polymerase chain reaction-based methodologies for gene doping detection are targeting exon-exon junctions in the intron-less transgene. However, because these junctions are known, it would be relatively easy to evade detection by tampering with the copyDNA sequences. We have developed a targeted next-generation sequencing based assay for the detection of all exon-exon junctions of the potential doping genes, EPO, IGF1, IGF2, GH1, and GH2, which is resistant to tampering. Using this assay, all exon-exon junctions of copyDNA of doping genes could be detected with a sensitivity of 1296 copyDNA copies in 1000 ng of genomic DNA. In addition, promotor regions and plasmid-derived sequences are readily detectable in our sequence data. While we show the reliability of our method for a selection of genes, expanding the panel to detect other genes would be straightforward. As we were able to detect plasmid-derived sequences, we expect that genes with manipulated junctions, promotor regions, and plasmid or virus-derived sequences will also be readily detected.

KW - gene doping

KW - next generation sequencing

U2 - 10.1038/s41434-019-0091-6

DO - 10.1038/s41434-019-0091-6

M3 - Article

VL - 26

SP - 338

EP - 346

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 7-8

ER -

ID: 90547137