Establishment of MNSs haplotyping methods by gene analytical techniques.

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(1)Investigations on genotypilg MNSs blood group Simple and accurate methods to genotype MN and Ss blood group systems by PCR were investigated, and the methods for MN genotyping using restriction fragment length polymorphism or single strand conformation polymorphism (SSCP) analysis, and the method for Ss genotyping using allele-specific PCR amplification were established and reported. Experimental condition for SSCP analysis was then studied and reported as a research manuscript.Several DNA samples which may involve duplicated alleles of MN blood group gene were found during the studies, and other polymorphic sites in the gene have been searched to analyze the duplicated allele.(2)Exploration and investigation of haplotyping methodsDuring the studies described above, conventional M allele of MN blood group gene was found to be divided into two sub-alleles, MG and MT, and their allele frequencies in Japanese, northern and southern Chinese populations were evaluated. To establish the method to analyze haplotypes consisting of different polymorphic genes such as MN and Ss genes, a simple allele-specific inverse PCR technique was applied first to detect an intragene haplotype of MN system consisting of MG, MT, and N.This technique (inverse PCR technique) was applied to detect an intragene haplotype of ABO blood group system, in which distance between polymorphic sites in ABO gene was much greater than that in MN gene.Further investigation is being continued to establish the method to analyze MNSs (intergene) haplotype.