Over recent years, requests for mtDNA
analysis in the field of forensic medicine
have notably increased, and the results of
such analyses have proved to be very useful
in forensic cases where nuclear DNA analysis cannot be performed. Traditionally,
mtDNA has been analyzed by DNA sequencing of the two hypervariable regions,
HVI and HVII, in the D-loop. DNA sequence analysis using the conventional
Sanger sequencing is very robust but time
consuming and labor intensive. By contrast,
mtDNA analysis based on the pyrosequencing technology provides fast and accurate
results from the human mtDNA present in
many types of evidence materials in forensic
casework. The assay has been developed to
determine polymorphic sites in the mitochondrial D-loop as well as the coding region to further increase the discrimination
power of mtDNA analysis. The pyrosequencing technology for analysis of mtDNA
polymorphisms has been tested with regard
to sensitivity, reproducibility, and success
rate when applied to control samples and
actual casework materials. The results
show that the method is very accurate and
sensitive; the results are easily interpreted
and provide a high success rate on casework samples. The panel of pyrosequencing
reactions for the mtDNA polymorphisms
were chosen to result in an optimal discrim ination power in relation to the number of
bases determined.