Determination of ABO genotypes with DNA extracted from formalin-fixed, paraffin-embedded tissues.
The gene encoding the specific glycosyltransferases which catalyze the conversion of the H antigen to A or B antigens shows a slight but distinct variation in its allelic nucleotide sequence and can be divided into 6 genotypes when digested with specific restriction enzymes. We extracted DNA from formalin-fixed, paraffin-embedded tissues using SDS/proteinase K treatment followed by phenol/chloroform extraction. The sequence of nucleotides for the A, B and O genes was amplified by the polymerase chain reaction (PCR). DNA fragments of 128 bp and 200 bp could be amplified in the second round of PCR, using an aliquot of the first round PCR product as template. Degraded DNA from paraffin blocks stored for up to 10.7 years could be successfully typed. The ABO genotype was deduced from the digestion patterns with an appropriate combination of restriction enzymes and was compatible with the phenotype obtained from the blood sample.
Yamada
M
M
Department of Legal Medicine, Shiga University of Medical Science, Otsu, Japan.
Yamamoto
Y
Y
Tanegashima
A
A
Kane
M
M
Ikehara
Y
Y
Fukunaga
T
T
Nishi
K
K
D000017:ABO Blood-Group System / Q000235:genetics*; D001483:Base Sequence; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D005554:Forensic Medicine / Q000379:methods; D005557:Formaldehyde; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA / Q000379:methods*; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods*
D000017:ABO Blood-Group System; D005557:Formaldehyde