|Budget Amount *help
¥7,700,000 (Direct Cost : ¥7,700,000)
Fiscal Year 1997 : ¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1996 : ¥5,600,000 (Direct Cost : ¥5,600,000)
HLA typing of both donor and recipients in organ transplantation is important from the stand point of equity and efficiency in unrelated cadaver organ transplantation. We have already developed a new HLA-DNA typing method using a RNA amplification method targeting at the genomic DNA,Transcription-Mediated Amplification (TMA) and a chemiluminescence-based Hybridization Protection Assay (HPA), which is one type of sequence-specific oligonucleotide (SSO) methods. In this research, we combined polymerase chain reaction (PCR) with HPA method, to analyze further fine alleles of the DR6 antigen group. One-hundred and twenty-one DNA samples were amplified with DR356-specific primers by PCR and detected by HPA method. Among DRB1^<**>13 allele group, 10 alleles (DRB1^<**>130l-1307, ^<**>1310-1312) were derfinitely typed, and among DRB1^<**>14,11 a11eles (DRBl^<**>l401-1408, ^<**>1411^<, **>1412, ^<**>l417). However, a certain combination of alleles (^<**>1301/02, ^<**>1140l/07) were not typed unequivocally and need additional primer combinations. The procedure time required for PCR-HPA method is one hour for extraction of DNA,2 hours for amplification, and one hour for detection, a total of 4 hours. Thus, PCR-HPA method for HLA-DNA typing is a simple, fast, non-isotopic and accurate method, which will guarantee the equity and efficiency in organ transplantation.