Genotyping of 19 red cell antigens, including RHD, using liquid bead arrays.
BACKGROUND
Traditionally, the determination of blood group has been performed using serological techniques. Due to the drawbacks of using antisera, molecular typing of blood groups has been introduced. However, the commonly used genotyping assays in blood banks have limitations because the panels for these were based on genetic variations in Caucasians.


METHODS
We developed a multiplex human erythrocyte antigen genotyping assay using allele-specific primer extensions and hybridization with bead microarrays. Single nucleotide polymorphisms (SNP) were genotyped for the 18 red cell antigens and wild-type RHD, DEL, and complete deletion were examined for RHD. The cut-off of median fluorescence intensity (MFI) for each allele was optimized. In the evaluation stage, the results of 87 samples were compared with those from other genotyping methods, and 26 serologic results were also compared.


RESULTS
The cut-off values were determined using -1 and -2 standard deviation (SD) of the minimum adjusted MFI for SNP detection. Complete deletion was determined with raw MFI+2SD. In comparison with the other methods, the kappa values were 0.984 overall. Compared with serologic methods, our assay showed discrepancy in 2 S/s, 3 C/c, and 3 Dia/Dibantigens.


CONCLUSIONS
Our method reliably predicts the presence or the absence of the 19 antigens.
Department of Laboratory Medicine, Sheikh Khalifa Specialty Hospital, Ras Al Khaimah, United Arab Emirates; Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Molecular Biology, Dankook University, Yongin, Republic of Korea.
Ho Eun
HE
Chang
Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Yun Ji
YJ
Hong
Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Hyungsuk
H
Kim
Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Sang Mee
SM
Hwang
Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Jeong Su
JS
Park
Department of Molecular Biology, Dankook University, Yongin, Republic of Korea.
Seong-Wook
SW
Lee
Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Eun Young
EY
Song
Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: m91w95pf@snu.ac.kr.
Kyoung Un
KU
Park
Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Junghan
J
Song
Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Kyou-Sup
KS
Han
D000483:Alleles; D001789:Blood Group Antigens / Q000097:blood / Q000235:genetics*; D005838:Genotype; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
D001789:Blood Group Antigens
Diego; Liquid bead; Luminex; Multiplex HEA genotyping; RHD