title: A rapid and accurate approach to identify single nucleotide polymorphisms of mitochondrial DNA using MALDI-TOF mass spectrometry.
abstract: 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.
authors.affiliation: Department of Research, University of Basel, Basel, Switzerland.
authors.forename: Alex
authors.initials: A
authors.lastname: Xiu-Cheng Fan
authors.forename: Henk S P
authors.initials: HS
authors.lastname: Garritsen
authors.forename: Shereen E L
authors.initials: SE
authors.lastname: Tarhouny
authors.forename: Michael
authors.initials: M
authors.lastname: Morris
authors.forename: Sinuhe
authors.initials: S
authors.lastname: Hahn
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Holzgreve
authors.forename: Xiao Yan
authors.initials: XY
authors.lastname: Zhong
mesh_terms: D001781:Blood Component Removal; D001792:Blood Platelets / Q000737:chemistry; D002467:Cell Nucleus / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics*; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
chemical_list: D004272:DNA, Mitochondrial