Korean J Blood Transfus.  2014 Dec;25(3):249-259. 10.0000/kjbt.2014.25.3.249.

Genotyping for RhD and RhCEce Antigens Using Free Circulating Nucleic Acids in Plasma and Serum

• Chang HE
• Hwang SM
• Hong YJ
• Han M
• Park JS
• Park KU
• Song J
• Han KS

• KMID: 2090035
• DOI: http://doi.org/10.0000/kjbt.2014.25.3.249

Abstract

BACKGROUND
The Rh blood group includes several antigens, of which D, C, E, c, and e are clinically important. Although nucleic acids from whole blood can be used for Rh blood group genotyping, it is also possible to genotype free circulating fetal nucleic acids from plasma and serum. We performed Rh blood group phenotyping and genotyping using nucleic acids from whole blood and free circulating nucleic acids from plasma and serum, respectively. The results were compared.
METHODS
Forty-four blood samples were phenotyped and genotyped for RhD and RhCE blood groups. Phenotyping was performed by hemagglutination assay. Further tests were performed on RhD-negative samples. Nucleic acids were extracted from whole blood, plasma, and serum. Plasma and serum were prepared after filtration and genotyped by real-time polymerase chain reaction.
RESULTS
RhD blood group results showed one (2.3%) discrepant case in which the DEL phenotype appeared wild RHD genotype. Among nucleic acids, there were seven discrepant results: two from plasma and five from serum based on whole blood nucleic acids. RhCE blood group results showed three (6.8%) phenotype-genotype discordances. Among nucleic acids, seven (15.9%mpared to phenotypes. Kappa coefficients of serum were lower than those of plasma.
CONCLUSION
RHD and RHCE genotype could be identified by assaying free circulating nucleic acids in plasma or serum. This study suggests that plasma is more reliable than serum as a specimen for RHD and RHCE genotyping of free circulating nucleic acids.

Keyword

RHD; RHCE; Genotyping; Free circulating nucleic acids

MeSH Terms

Blood Group Antigens
Filtration
Genotype
Hemagglutination
Nucleic Acids*
Phenotype
Plasma*
Real-Time Polymerase Chain Reaction
Blood Group Antigens
Nucleic Acids