Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jun 25;10(13):e33608.
doi: 10.1016/j.heliyon.2024.e33608. eCollection 2024 Jul 15.

Simultaneous high throughput genotyping of 36 blood group systems using NGS based on probe capture technology

Jingjing Zhang  1 , Wenjing Yuan  1 , Xiaozhen Hong  1 , Yanling Ying  1 , Faming Zhu  1
Affiliations

Simultaneous high throughput genotyping of 36 blood group systems using NGS based on probe capture technology

Jingjing Zhang et al. Heliyon. .

Abstract

Human blood group antigen has important biological functions, and transfusion of incompatible blood can cause alloimmunization and may lead to serious hemolytic reactions. Currently, serological methods are most commonly used in blood group typing. However, this technique has certain limitations and cannot fully meet the increasing demand for the identification of blood group antigens. This study describes a next-generation sequencing (NGS) technology platform based on exon and flanking region capture probes to detect full coding exon and flanking intron regions of the 36 blood group systems, providing a new high-throughput method for the identification of blood group antigens. The 871 capture probes were designed for the exon and flanking intron sequences of 36 blood group system genes, and synchronization analysis for 36 blood groups was developed. The library for NGS was tested using the MiSeq Sequencing Reagent Kit (v2, 300 cycles) by Illumina NovaSeq, and the data were analyzed by the CLC Genomics Workbench 21.0 software. A total of 199 blood specimens have been sequenced for the 41 genes from 36 blood groups. Among them, heterozygote genotypes were found in the ABO, Rh, MNS, Lewis, Duffy, Kidd, Diego, Gerbich, Dombrock, Globoside, JR, LAN, and Landsteiner-Wiene blood group systems. Only the homozygous genotype was found in the remaining 22 blood group systems. The obtained data in the NGS method shows a good correlation (99.98 %) with those of the polymerase chain reaction-sequence-based typing. An NGS technology platform for 36 blood group systems genotyping was successfully established, which has the characteristics of high accuracy, high throughput, and wide coverage.

Keywords: Blood group genotyping; Exon and flanking intron sequences; NGS; Probe capture technology.

PubMed Disclaimer

Conflict of interest statement

No conflict of interest exits in the submission of this manuscript, and manuscript is approved by all authors for publication.

Figures

Fig. 1
Fig. 1
Results of NGS genotyping of ABO blood group in 199 blood donors. Different colors indicate different genotypes. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 2
Fig. 2
CLC Genomics Workbench 21.0 software data analysis steps.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Mohandas N., Narla A. Blood group antigens in health and disease. Curr. Opin. Hematol. 2005;12(2):135–140. doi: 10.1097/01.moh.0000153000.09585.79. - DOI - PubMed
    1. Tsintsadze I., Akhvlediani L., Khukhunaishvili R., Koridze M., Tskvitinidze S., NakaShidze I., Dolidze K., Loria R., Nagervadze M. Specific combinations of erythrocyte group antigens in blood donors. Indian journal of hematology & blood transfusion : an official journal of Indian Society of Hematology and Blood Transfusion. 2020;36(2):356–360. doi: 10.1007/s12288-019-01197-4. - DOI - PMC - PubMed
    1. Krog G.R., Rieneck K., Clausen F.B., Steffensen R., Dziegiel M.H. Blood group genotyping of blood donors: validation of a highly accurate routine method. Transfusion. 2019;59(10):3264–3274. doi: 10.1111/trf.15474. - DOI - PubMed
    1. Moncharmont P. Red blood cell alloimmunisation after platelet transfusion (excluding ABO blood group system) Transfus. Clin. Biol. : journal de la Societe francaise de transfusion sanguine. 2020;27(3):185–190. doi: 10.1016/j.tracli.2020.06.001. - DOI - PubMed
    1. Tormey C.A., Hendrickson J.E. Transfusion-related red blood cell alloantibodies: induction and consequences. Blood. 2019;133(17):1821–1830. doi: 10.1182/blood-2018-08-833962. - DOI - PMC - PubMed

LinkOut - more resources