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Genotyping Parkinson Disease-Associated Mitochondrial Polymorphisms



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  • Personal Author:
  • Description:
    OBJECTIVE: The purpose of this study was to establish a system for rapidly detecting single nucleotide polymorphisms (SNPs) in mitochondrial DNA (mtDNA) using hybridization probes and melting temperature (T(m)) analysis. This technology should prove useful for population-based studies on the interaction between genetic factors and environmental exposures and the risk of Parkinson disease (PD). METHODS: Mitochondrial DNA (mtDNA) was extracted from whole blood. Rapid polymerase chain reaction (PCR) and melting curve analyses were performed with primers and fluorochrome-labeled probes on a LightCycler (Roche Molecular Biochemical, Mannheim, Germany). Genotyping of 10 SNPs in 15 subjects was based on the analysis of allele-specific T(m) of detection probes. The results of melting curve analyses were verified by sequencing all 150 PCR products. RESULTS: Real-time monitoring showed optimal PCR amplification of each mtDNA fragment. The nucleotide changes at positions 1719, 4580, 7028, 8251, 9055, 10398, 12308, 13368, 13708, and 16391 from wild-type to mutant genotype resulted in 6.51, 8.29, 3.26, 7.82, 4.79, 2.84, 2.73, 9.04, 8.53, and 9.52 degrees C declines in T(m) of the detection probes, respectively. Genotyping of all 150 samples was verified by 100% correspondence with the results of sequencing. Fourteen subjects were haplogrouped by combining results for all 10 SNPs. CONCLUSION: A rapid and reliable detection system for identifying mitochondrial polymorphisms and haplotypes was developed based on hybridization probe technology. This method may be suitable for mitochondrial genotyping of samples from large-scale epidemiology studies, and may prove useful for exploring the molecular etiopathogenesis of PD, identifying markers of genetic susceptibility, and protecting susceptible individuals from PD. [Description provided by NIOSH]
  • Subjects:
  • Keywords:
  • ISSN:
    1539-4182
  • Document Type:
    Text
  • Funding:
    Cooperative Agreement
  • Genre:
    Journal Article
  • Place as Subject:
  • CIO:
    National Institute for Occupational Safety and Health (NIOSH)
  • Topic:
    Workplace Safety & Health
  • Location:
    North America
  • Pages in Document:
    99-106
  • Volume:
    2
  • Issue:
    2
  • NIOSHTIC Number:
    nn:20029118
  • Citation:
    Clin Med Res 2004 May; 2(2):99-106
  • Contact Point Address:
    National Farm Medicine Center, Marshfield Clinic Research Foundation, Marshfield, Wisconsin 54449
  • Federal Fiscal Year:
    2004
  • Performing Organization:
    Marshfield Clinic Research Foundation
  • Peer Reviewed:
    True
  • Start Date:
    20010930
  • Source Full Name:
    Clinical Medicine & Research
  • End Date:
    20040929
  • Collection(s):
  • Main Document Checksum:
    urn:sha-512:776d51270d3dc5fc98124974ed11bfd466ca81746239c9ef434a9b36bbd39a3e6801db8d4cc14f0cb5f1b286fcf35fe4aa6c071903884a6ca5f5ff7731bae01a
  • Download URL:
  • File Type:
    Filetype[PDF - 499.19 KB ]
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