Because of the sensitivity of detection of mitochondrial DNA (mtDNA) analysis, a higher success rate from old samples, severely decomposed or charred remains is achieved with mtDNA compared with nuclear DNA. Since mtDNA is maternally inherited, the sequence analysis of human mtDNA is being used widely for evidence of mother and child relationship or a sibling case. However, the mtDNA has a higher mutation rate than the nuclear DNA. Some regions of the mtDNA appear to be nucleotide substitution in all maternally linked relatives. in this study, we have carried out simultaneous typing to 11 SNPs and 2 tandem repeat sites in the mtDNA by use of the allele-specific amplification technique from 100 mother-child pairs and 60 sibling pairs. As a result, we found 9 sample pairs, out of 160 sample pairs, which appeared to be of different band intensity of each other. In a compared analysis to the 9 sample pairs, we have concluded that the difference of band intensity is attributable to point and length heteroplasmy of the mtDNA.