Development and verification of PCR-fingerprinting HLA-DPB1 genotyping
- VernacularTitle:聚合酶链反应指纹图HLA-DPB1分型法的建立与验证
- Author:
Ming WU
;
Lianhuang LU
- Publication Type:Journal Article
- Keywords:
PCR-fingerprinting DNA HLA antigens
- From:
Chinese Journal of Organ Transplantation
1998;19(3):133-136,封3
- CountryChina
- Language:Chinese
-
Abstract:
To accelerate the identification of HLA-DPB1 matched marrow donors from unrelated population, a very simple HLA-DPB1 genotyping method called PCR-fingerprinting (PCRF) was developed according to the theory about homoduplex and heteroduplex formation from different PCR coding strands and non-coding ones. Unlike PCR-SSCP, strict laboratory condition is not needed in the PCRF. After denaturing at 94℃ for 2 min and cooling at 37℃ for 8 min, the PCR product was separated by 8% PAGE for 5 h and polymorphism band patterns would appear when the gel staining was completed with either EB or silver staining procedure. To confirm its reliability, 21 individuals from 9 family whose DPB1 genotypes assigned by PCR-RFLP were verified. It was found that there were 8 PCRF patterns corresponding to the 9 HLA-DPB1 genotypes from the 21 cases and the same DPB1 genotypes produced identical PCRF pattern except one pair.The factors on efficient separation of heteroduplexes and homoduplexes were also discussed.
