Application of third-generation sequencing in monitoring the proportion of donor gene chimerism after allogeneic hematopoietic stem cell transplantation in patients with beta-thalassemia major
10.3760/cma.j.cn114452-20240525-00271
- VernacularTitle:第三代测序在重型β地中海贫血患者allo-HSCT术后供体基因嵌合比例监测中的应用研究
- Author:
Linlin LI
1
;
Yifang HUANG
;
Yunhua HUANG
;
Liqiu PAN
;
Zuhao WU
;
Faquan LIN
Author Information
1. 广西医科大学第一附属医院检验科 广西高校临床检验诊断学重点实验室，南宁 530021
- Keywords:
beta-Thalassemia;
Hematopoietic stem cell transplantation;
Third generation sequencing;
Chimerism
- From:
Chinese Journal of Laboratory Medicine
2024;47(9):1059-1066
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To monitor the changes in donor gene chimerism ratio after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with severe beta-thalassemia using third-generation sequencing, and to explore the value of this technology in monitoring the proportion of donor genes chimerism in the early stage of postoperative allo-HSCT.Methods:Case analysis. Three beta-thalassemia patients at the First Affiliated Hospital of Guangxi Medical University during June-July 2022 who had undergone allo-HSCT with genotypes IVS-Ⅱ-654/CD41-42, IVS-Ⅱ-654/IVS-Ⅱ-654 and CD41-42/CD41-42 were included in this study. "Visual" analysis of the readouts of recipient DNA using third generation sequencing was used to monitor the genetic chimerism of the donor DNA and to compare with Sanger sequencing results. Post-transplantation follow-up was performed in the three patients to monitor the blood statistics and assess their implantation status and hematopoietic reconstitution.Results:The results of donor DNA chimerism status after allo-HSCT in the three patients detected by third generation sequencing were consistent with the Sanger sequencing results. The chimeric state of donor DNA gradually shifted to complete donor gene chimerism as the number of days after transplantation increased. Recipient 1 had 95.5% and 100% donor DNA chimerism at 10 and 20 d post-transplantation, respectively; recipient 2 had 100% donor DNA chimerism at 30 and 40 d post-transplantation; recipient 3 had 69.5% donor DNA chimerism at 1 d post-transplantation, and 100% donor DNA chimerism at 10 and 20 d post-transplantation. All patients achieved full donor gene chimerism within 30 d post-transplantation. Stable implantation of granulopoiesis, platelets, and erythropoiesis with hematopoietic reconstitution were obtained in all 3 patients within 1 month after transplantation.Conclusions:In this study, we developed a new method to detect the chimerism ratio of donor DNA using third-generation sequencing technology, enabling us to monitor the gene chimerism status of donor DNA at an early stage.
