| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Research Abstract |
We studied the genotping of the ABO blood group system by using the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods. The DNA fragments containing the nucleotide (nt) position 261,467,526,646,703,796,802 and 803 of cDNA from A-transferase were amplified by PCR with the primers designed in the present study. The results obtained are as follows. 1. The eight nucleotide positions were analyzed by PCR-RFLP and allele specific PCR (asPCR). The ABO genotyping was carried out successfully, and common A alleles were divided into two suballeles named A (Pro) and A (Leu), and common 0 alleles were divided into 0(T) and 0(A) suballeles. No suballele of B was found. 2. From the result of nt802 analysis, we found a variant allele, 0^2, in German samples (allele frequency : >2%). The 0^2 allele has not been found in Japanese samples so far. 3. A case of blood chimera was found in a Japanese family. The family consisted of both parents and three children. The results from the conventional serological typings showed that the mother was not the 'biological mother' of the children. Then, we performed the genotyping of the ABO blood group system by using PCR-RFLP and asPCR.The asPCR of the mother's DNA amplified A-allele-, B-allele- and O-allele-specific DNAs, showing that she had a very rare type, blood group chimera. The ABO genotype of the mother was A(Leu)/B/O(T), and the finding made it manifest that the mother was the biological mother of the children. We got many good results from this research as expected.
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