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Development of a Rapid, Immobilized Probe Assay for the Detection of Mitochondrial DNA Variation in the HVI and HVII Regions

NCJ Number
Date Published
49 pages
This report describes the development of a rapid amplification and typing system for the analysis of Mitochondrial DNA sequence variation in the HVI and HVII regions.
In 1981 Anderson et al. first sequenced the human mitochondrial genome and since that time, the sequence diversity of the mitochondrial genome has been well documented. This report focuses on the development of a sensitive and robust amplification and typing system that will allow for the detection of most of the diversity in the HVI and HVII regions of the human mitochondrial genome. The new system is more cost and time effective than previous methods. The initial version of the system adapted 23 Sequence-Specific Oligonucleotide (SSO) probes that used a standard dot blot format to the immobilized probe method. Following the initial version, the first linear array probe panel for mtDNA sequence analysis included 17 SSO probes targeting the HVII region. In order to increase the flexibility and discrimination of the assay, a primer pair for the HVI region was incorporated into the PCR amplification reaction and 18 additional probes were added to detect sequence variation in 4 HVI regions and 2 heteroplasmy hotspots. The development validation of the resulting LINEAR ARRAY mtDNA HVI/HVII Region-Sequence Typing Kit has been completed and internal validation by DNA testing laboratories is ongoing. Additionally, the assay has been successfully used in Sweden for the analysis of over 300 forensic samples and for the identification of mass grave remains in Croatia. Tables, figure, references

Date Published: January 1, 2005