Genotyping of the ABCG2 gene using Matrix‐Associated Laser Desorption/Ionisation, Time‐of‐Flight Mass Spectrometry

• Published in: Transfusion medicine (Oxford, England) Vol. 28; no. 3; pp. 255 - 260
• Main Authors: Tanaka, M., Kamada, I., Takahashi, J., Kimura, T., Tani, Y.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.06.2018
• Subjects: ABCG2 gene; Asian Continental Ancestry Group; ATP Binding Cassette Transporter, Sub-Family G, Member 2 - genetics; DNA‐based genotyping; Female; Genotyping Techniques - instrumentation; Genotyping Techniques - methods; Humans; Japan; Jr (a−) phenotype; MALDI –TOF; Male; Neoplasm Proteins - genetics; Polymorphism, Single Nucleotide; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; ABCG2 gene; MALDI -TOF; Jr (a−) phenotype; DNA-based genotyping
• ISSN: 0958-7578; 1365-3148
• DOI: 10.1111/tme.12474

Objectives The aim of this study was to evaluate the applicability of genotyping of the ABCG2 gene using MALDI‐TOF MS and to estimate the allele frequency in the Japanese population. Background Jr (a−) phenotype has a prevalence of approximately 0·05% among Japanese blood donors; DNA‐based genotyping was conducted to investigate the molecular basis of the Jr (a−) phenotype along with serological typing. To detect all SNPs of the ABCG2 gene, a high‐throughput SNP genotyping platform is needed. Methods Overall, 1004 Jr (a−) blood samples were collected from blood donors in Japan and pre‐genotyped. To detect the SNPs of the ABCG2 gene using MALDI‐TOF MS, polymerase chain reaction and unextend primer were designed. In total, 205 Jr (a−) samples were genotyped using MALDI‐TOF MS analysis. Results The SNPs of 1004 Jr (a−) samples were identified using the HRM analysis and DNA sequencing, and 799 of 1004 (80%) Jr (a−) samples had the homozygous for c.376 T. The designed primers for MALDI‐TOF MS perfectly detected the SNPs of the ABCG2 gene. A total of 205 Jr (a−) samples were genotyped using MALDI‐TOF MS. Calling failures occurred in only two samples with the mutations c.736CT to c.376C and c.421C to c.421CA. The concordance rate between the pre‐genotyped and MALDI‐TOF MS‐based genotyping results was very high (99·02%) for all ABCG2 alleles. Conclusions Jr (a‐) Japanese donors had almost the homozygous for c.376 T. However, detections of more than 20 SNPs of the ABCG2 gene for the JR blood group genotyping are needed. MALDI‐TOF MS‐based genotyping was highly concordant with the pre‐genotyped results for all ABCG2 alleles.