[ABO genotyping by PCR-direct sequencing method].
OBJECTIVE
To analyze the sequence difference between human A, B, and O alleles and establish the method of ABO genotyping by PCR direct sequencing.


METHODS
PCR-direct sequencing technique was used to analyze two regions of cDNA from A transferase gene, 233-433 and 660-788.


RESULTS
Two nucleotide substitutions at 258th and 297th were found in 233-433 region, and a nucleotide substitution at 700th was found in 660-788 region. At 258th, the nucleotide was guanine in A and B alleles, and adenine in O allele. At 297th, the nucleotide was adenine in A allele, and guanine in B allele. As this position, O allele was subdivided into two types, O(A) and O(G). At 700th, the nucleotide was guanine in A and O alleles, and adenine in B allele. Therefore, 8 genotypes, AA, AO(A), AB, BB, BO(G), O(A) O(A), O(G) O(G) and O(A) O(G), could be clearly determined by only analyzing the 233-433 region. The other two genotypes, AO(G) and BO(A), could be further distinguished by analyzing the 660-788 region.


CONCLUSIONS
The technique of PCR-direct sequencing provides an effective and new method for ABO genotyping further.
Jiang
X
X
Liaoning Province Criminal Science and Technology Institute, Shenyang, Liaoning, 110032 P.R.China.
Hou
G
G
Yu
J
J
Huang
B
B
Xu
D
D
D000017:ABO Blood-Group System / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000017:ABO Blood-Group System; D018076:DNA, Complementary