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

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Immunohematology

American National Red Cross

Subject: Medical Laboratory Technology

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ISSN: 0894-203X
eISSN: 1930-3955

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VOLUME 30 , ISSUE 4 (December 2014) > List of articles

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

Tianxiang Gong / Ying Hong / Naihong Wang / Xuemei Fu / Changhua Zhou

Keywords : blood group genotyping, real-time PCR, melting curve analysis

Citation Information : Immunohematology. Volume 30, Issue 4, Pages 161-165, DOI: https://doi.org/10.21307/immunohematology-2019-114

License : (Transfer of Copyright)

Published Online: 01-December-2019

ARTICLE

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 realtime 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.

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