HLA-B60 and HLA-B61 Discrimination by PCR using Sequence-specific Primers (PCR-SSP) Method.
- Author:
Hee Yeon WOO
1
;
Yun Sun YANG
Author Information
1. Department of Clinical Pathology, Sungkyunkwan University School of Medicine, Seoul, Korea
- Publication Type:Original Article
- Keywords:
HLA-B40;
B60;
B61;
PCR-SSP
- MeSH:
Alleles;
Discrimination (Psychology)*;
Exons;
Fetal Blood;
Heterozygote;
HLA-B Antigens;
HLA-B40 Antigen;
Homozygote;
Humans;
Indicators and Reagents;
Lymphocytes;
Molecular Typing;
Organ Transplantation;
Polymerase Chain Reaction*;
Sensitivity and Specificity;
Tissue Donors;
Transplants
- From:Korean Journal of Clinical Pathology
1999;19(6):702-706
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: HLA-B40 is the most frequently identified HLA-B type in Koreans. Also HLA-B60 and B61 are the serologic split antigens of HLA-B40. But because of the lack of mono-specific alloantisera, cross reactivity of sera used as typing reagents, and poor antigenicity of some specific cells such as cord blood lymphocytes, discrimination between HLA-B60 and B61 has been often problematic in laboratories. In this study, authors evaluated whether the PCR-SSP method can be useful for accurate assignments of HLA-B60 and B61 or not. METHODS: Twenty-nine lymphocytes samples which were suspected as heterozygotes or homozygotes of HLA-B60 or B61 and three samples typed as HLA-B40 are selected from stored cord blood and organ transplantation donors. HLA types of these samples were defined by serologic method using a commercial typing kit. PCR that amplified exons 2 and 3 of the HLA-B gene using sequence specific primer pairs exactly matched to HLA-B60 or B61 allele making up a serological specificity was done. RESULTS: A clear discrimination between B60 and B61 was possible in all samples including 9 serologically ambiguous samples. Discrepancy between serologic typing and molecular typing was seen in three cases identified serologically as B40 positive but inable to define a split. Among three samples, two were identified as HLA-B61 and one was identified as HLA-B60. CONCLUSIONS: Molecular typing was useful in discriminating between HLA-B60 and B61. The PCR-SSP method for HLA-B60 and B61 including other cross-reactive HLA types will be helpful as a supplemental method of the serologic typing.
