[PCR/SSCP used as a complementary method for HLA oligotyping].
Seventy samples randomly selected from Hunan Han nationality normal individuals were typed for HLA DPA1 polymorphism by PCR/SSO with the primers and SSOs. Silver staining PCR/SSCP was also performed on the polymorphic second exon of all samples, which demonstrated the accurate gene frequencies of DPA1*01,02 to be 0.2632 and 0.7327, respectively. Using PCR/SSCP technique, we found that PCR/SSO mistyped a DPA1*01/02 heterozygote as single DPA1*01 gene because of weak hybridization signals. PCR/SSCP also revealed DPA1*02 in Hunan Han population to be composed of a single subtype. The results indicate that PCR/SSCP can be used to clarify the samples with atypical or irregular PCR/SSO hybridization patterns and to analyze the possible polymorphisms outside SSOs sequences. Thus this method may guarantee the accuracy of HLA oligotyping.
Tian
W
W
Department of Immunology, Hunan Medical University, Changsha 410078.
Guo
S
S
Li
L
L
D005787:Gene Frequency; D006682:HLA-DP Antigens / Q000145:classification / Q000235:genetics*; D059867:HLA-DP alpha-Chains; D006801:Humans; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational
D006682:HLA-DP Antigens; D059867:HLA-DP alpha-Chains; C071172:HLA-DPA1 antigen; D009841:Oligonucleotides