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A rapid and accurate approach to identify single nucleotide polymorphisms of mitochondrial DNA using MALDI-TOF mass spectrometry

Xiu-Cheng Fan, Alex ; Garritsen, Henk S.P. ; Tarhouny, Shereen E.L. ; Morris, Michael ; Hahn, Sinuhe ; Holzgreve, Wolfgang ; Zhong, Xiao Yan

In: Clinical Chemical Laboratory Medicine, 2008, vol. 46, no. 3, p. 299-305

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    Titre
    • A rapid and accurate approach to identify single nucleotide polymorphisms of mitochondrial DNA using MALDI-TOF mass spectrometry
    Auteur
    • Xiu-Cheng Fan, Alex. The first two authors contributed equally to this work.Laboratory for Prenatal Medicine and Gynecologic Oncology, Women's Hospital/Department of Research, University of Basel, Basel, Switzerland
    • Garritsen, Henk S.P.. Institute for Clinical Transfusion Medicine, Staedtisches Klinikum Braunschweig GmbH, Braunschweig, Germany
    • Tarhouny, Shereen E.L.. Laboratory for Prenatal Medicine and Gynecologic Oncology, Women's Hospital/Department of Research, University of Basel, Basel, Switzerland and Medical Biochemistry Department, Faculty of Medicine, Zagazig University, Zagazig, Egypt
    • Morris, Michael. Laboratoire de Diagnostic Moleculaire, Service de Medicine Genetique, Geneva, Switzerland
    • Hahn, Sinuhe. Laboratory for Prenatal Medicine and Gynecologic Oncology, Women's Hospital/Department of Research, University of Basel, Basel, Switzerland
    • Holzgreve, Wolfgang. Laboratory for Prenatal Medicine and Gynecologic Oncology, Women's Hospital/Department of Research, University of Basel, Basel, Switzerland
    • Zhong, Xiao Yan. Laboratory for Prenatal Medicine and Gynecologic Oncology, Women's Hospital/Department of Research, University of Basel, Basel, Switzerland
    Type de document
    • Postprint
    Langue
    • Inglese
    Publié dans
    • Clinical Chemical Laboratory Medicine, 2008, vol. 46, no. 3, p. 299-305. Walter de Gruyter
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:300936
    Summary
    Background: Single nucleotide polymorphisms (SNPs) of mitochondrial DNA (mtDNA) are involved in physiological and pathological conditions. We developed a rapid, accurate, highly sensitive and high-throughput approach with low cost to identify mtDNA SNPs. Methods: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to detect 18 SNPs of mtDNA by uniplex and multiplex assays. The sensitivity and specificity of the MALDI-TOF MS were evaluated. The accuracy of the approach was validated by the comparison of using the robust sequencing analysis. Results: The detection limit achieved with the assays corresponded to the identification of five-genome equivalence of mtDNA per reaction after first round PCR amplification. The testing system enabled the discrimination of as little as 5% of mtDNA polymorphism in the predominating background of mtDNA not containing the SNP. No false positive and false negative results were obtained using the uniplex and multiplex MALDI-TOF MS assays for the analysis of the 18 SNPs compared with those obtained by sequencing analysis. Conclusions: Possible fields which could benefit from this powerful and sensitive tool include forensic medicine, tracing of matrilineage, transplantation immunology, transfusion medicine, the diagnosis of mtDNA mutation related disorders, and the research regarding aging, apoptosis and carcinogenesis based on physiologic and pathogenic alterations of mtDNA for the analysis of large-scale samples, multiple SNPs or rare mtDNA. Clin Chem Lab Med 2008;46:299-305