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Long-read sequencing on the SMRT platform enables efficient haplotype linkage analysis in preimplantation genetic testing for β-thalassemia

  • Genetics
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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

This study aimed to evaluate the value of long-read sequencing for preimplantation haplotype linkage analysis.

Methods

The genetic material of the three β-thalassemia mutation carrier couples was sequenced using single-molecule real-time sequencing in the 7.7-kb region of the HBB gene and a 7.4-kb region that partially overlapped with it to detect the presence of 17 common HBB gene mutations in the Chinese population and the haplotypes formed by the continuous array of single-nucleotide polymorphisms linked to these mutations. By using the same method to analyze multiple displacement amplification products of embryos from three families and comparing the results with those of the parents, it could be revealed whether the embryos carry disease-causing mutations without the need for a proband.

Results

The HBB gene mutations of the three couples were accurately detected, and the haplotype linked to the pathogenic site was successfully obtained without the need for a proband. A total of 68.75% (22/32) of embryos from the three families successfully underwent haplotype linkage analysis, and the results were consistent with the results of NGS-based mutation site detection.

Conclusion

This study supports long-read sequencing as a potential tool for preimplantation haplotype linkage analysis.

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Funding

This study received funding from sources as follows: the Natural Science Foundation of Guangdong Province, China (2018A030310050); National Key R&D Program of China (2016YFC1000205); Guangdong Provincial Key Laboratory of Reproductive Medicine (2012A061400003).

Author information

Author notes

  1. Haitao Wu and Dongjia Chen contributed equally to this work.

Authors and Affiliations

  1. Reproductive Medicine Center, Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-Sen University, Jiangmen, Guangdong, China

    Haitao Wu, Qiang Zhao, Yongbin Liao & Ping Li

  2. Reproductive Medicine Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China

    Dongjia Chen, Xiaoting Shen & Canquan Zhou

  3. Department of Obstetrics and Gynaecology, Queen Mary Hospital, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China

    Philip C. N. Chiu

  4. Shenzhen Key Laboratory of Fertility Regulation, Department of Obstetrics and Gynecology, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China

    Philip C. N. Chiu

Authors
  1. Haitao Wu
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  2. Dongjia Chen
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  3. Qiang Zhao
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  4. Xiaoting Shen
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  5. Yongbin Liao
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  6. Ping Li
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  7. Philip C. N. Chiu
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  8. Canquan Zhou
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Contributions

H.W., X.S., and C.Z. designed and performed the experiments, collected and analyzed data, and wrote the manuscript. D.C., H.W., Y.L., P.L., and Q.Z. conducted the experiment. D.C. contributed to the interpretation of the results. P.C.C. and C.Z. supervised the experiments, and revised the manuscript. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Philip C. N. Chiu or Canquan Zhou.

Ethics declarations

Ethics approval

This study was approved by the ethics committee of the Affiliated Jiangmen Hospital of Sun Yat-Sen University. We obtained informed consent from all the couples before this study.

Conflict of interest

The authors declare no competing interests.

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Wu, H., Chen, D., Zhao, Q. et al. Long-read sequencing on the SMRT platform enables efficient haplotype linkage analysis in preimplantation genetic testing for β-thalassemia. J Assist Reprod Genet 39, 739–746 (2022). https://doi.org/10.1007/s10815-022-02415-1

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  • DOI: https://doi.org/10.1007/s10815-022-02415-1

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