<abstract xmlns="http://www.w3.org/1999/xhtml"><p>The detection of polymorphism is the basis of blood group genotyping and phenotype prediction. Genotyping may be useful to determine blood groups when serologic results are unclear. The development and application of different methods for blood group genotyping may be needed as a substitute for blood group typing. The purpose of this study is to establish an approach for blood group genotyping based on a melting curve analysis of real-time polymerase chain reaction (PCR). Using DNA extracted from whole blood, we developed and validated a DNA typing method for detecting <italic>DO*01/DO*02, DI*01/DI*02, LU*01/LU*02,</italic> and <italic>GYPB*03/GYPB*04</italic> alleles using a melting curve analysis. All assays were confirmed with a commercial reagent containing sequence-specific primers (PCR-SSP), and a cohort of the samples was confirmed with sequencing. Results for all blood groups were within the range of specificity and assay variability. Genotypes of 300 blood donors were fully consistent with PCR-SSP data. The obtained genotype distribution is in complete concordance with existing data for the Chinese population. There are several advantages for this approach of blood group genotyping: lower contamination rates with PCR products in the laboratory, ease of performance, automation potential, and rapid cycling time. <italic><bold>Immunohematology</bold></italic><bold>2014;30:161–165.</bold></p></abstract>

The detection of polymorphism is the basis of blood group genotyping and phenotype prediction. Genotyping may be useful to determine blood groups when serologic results are unclear. The development and application of different methods for blood group genotyping may be needed as a substitute for blood group typing. The purpose of this study is to establish an approach for blood group genotyping based on a melting curve analysis of real-time polymerase chain reaction (PCR). Using DNA extracted from whole blood, we developed and validated a DNA typing method for detecting DO*01/DO*02, DI*01/DI*02, LU*01/LU*02, and GYPB*03/GYPB*04 alleles using a melting curve analysis. All assays were confirmed with a commercial reagent containing sequence-specific primers (PCR-SSP), and a cohort of the samples was confirmed with sequencing. Results for all blood groups were within the range of specificity and assay variability. Genotypes of 300 blood donors were fully consistent with PCR-SSP data. The obtained genotype distribution is in complete concordance with existing data for the Chinese population. There are several advantages for this approach of blood group genotyping: lower contamination rates with PCR products in the laboratory, ease of performance, automation potential, and rapid cycling time. Immunohematology2014;30:161–165.

Validation of a blood group genotyping method based on high-resolution melting curve analysis

Immunohematology

The detection of polymorphism is the basis of blood group genotyping and phenotype prediction. Genotyping may be useful to determine blood groups when...

Validation of a blood group genotyping method based on high-resolution melting curve analysis

Article Category: Report

Publié en ligne: 01 déc. 2019

Pages: 161 - 165

DOI: https://doi.org/10.21307/immunohematology-2019-114

Mots clés
blood group genotyping, real-time PCR, melting curve analysis

© 2014 T. Gong et al., published by Sciendo

Validation of a blood group genotyping method based on high-resolution melting curve analysis

Article Category: Report

Publié en ligne: 01 déc. 2019

Pages: 161 - 165

DOI: https://doi.org/10.21307/immunohematology-2019-114

Mots clésblood group genotyping, real-time PCR, melting curve analysis

© 2014 T. Gong et al., published by Sciendo

Mots clés
blood group genotyping, real-time PCR, melting curve analysis