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Validation of the multiplex ligation‐dependent probe amplification assay and its application on the distribution study of the major alleles of 17 blood group systems in C hinese donors from G uangzhou
Author(s) -
Ji Yanli,
Wen Jizhi,
Veldhuisen Barbera,
HaerWigman Lonneke,
Wang Zhen,
Lodénvan Straaten Martin,
Wei Ling,
Luo Guangping,
Fu Yongshui,
van der Schoot C. Ellen
Publication year - 2017
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.13940
Subject(s) - multiplex , allele , ligation , microbiology and biotechnology , biology , chemistry , medicine , genetics , gene
BACKGROUND Genotyping platforms for common red blood cell (RBC) antigens have been successfully applied in Caucasian and black populations but not in Chinese populations. In this study, a genotyping assay based on multiplex ligation‐dependent probe amplification (MLPA) technology was applied in a Chinese population to validate the MLPA probes. Subsequently, the comprehensive distribution of 17 blood group systems also was obtained. STUDY DESIGN AND METHODS DNA samples from 200 Chinese donors were extracted and genotyped using the blood‐MLPA assay. To confirm the MLPA results, a second independent genotyping assay (ID Core+) was conducted in 40 donors, and serological typing of 14 blood‐group antigens was performed in 91 donors. In donors who had abnormal copy numbers of an allele ( DI and GYPB ) determined by MLPA, additional experiments were performed (polymerase chain reaction, sequencing, and flow cytometry analysis). RESULTS The genotyping results obtained using the blood‐MLPA and ID Core+ assay s were consistent. Serological data were consistent with the genotyping results except for one donor who had a Lu(a−b−) phenotype. Of the 17 blood group systems, the distribution of the MNS, Duffy, Kidd, Diego, Yt, and Dombrock systems was polymorphic. The Mur and St a antigens of the MNS system were distributed with a frequency of 9% (18 of 200) and 2% (4 of 200), respectively. One donor with chimerism and one who carried a novel DI*02(A845V) allele, which predicts the depression of Di b antigen expression, were identified. CONCLUSIONS The blood‐MLPA assay could easily identify the common blood‐group alleles and correctly predicted phenotype in the Chinese population. The Mur and St a antigens were distributed with high frequency in a Southern Chinese Han population.