摘要
目的应用第三代测序技术监测重型β地中海贫血患者异基因造血干细胞移植(allo-HSCT)术后供体基因嵌合比例的变化,并初步探讨该技术在allo-HSCT术后早期供体基因嵌合比例监测中的应用价值。方法病例分析。以2022年6—7月在广西医科大学第一附属医院就诊的3例已行allo-HSCT的重型β地中海贫血患者为研究对象,基因型分别为IVS-Ⅱ-654/CD41-42、IVS-Ⅱ-654/IVS-Ⅱ-654和CD41-42/CD41-42。利用第三代测序技术“可视化”分析3例患者allo-HSCT术后供受体DNA的读长比例,从而监测供体DNA的基因嵌合状态,并与Sanger测序结果进行对比分析。对3例患者进行移植后随访,监测血象,评估其植入状态和造血重建情况。结果第三代测序检测的3例患者allo-HSCT术后供体DNA嵌合状态的结果与Sanger测序结果一致,随着移植后天数的增加,供体DNA的嵌合状态逐渐向完全供体基因嵌合状态转变。受者1移植后10 d、20 d供体DNA嵌合比例分别为95.5%和100%;受者2移植后30 d和40 d,供体DNA嵌合比例均达100%;受者3移植后1 d供体DNA嵌合比例为69.5%,移植后10 d、20 d均达100%;所有患者移植后30 d内均达到完全供体基因嵌合。3例患者均在移植后1个月内获得了粒系、血小板系、红系的稳定植入,造血重建。结论本研究开发了一种利用第三代测序技术检测供体DNA嵌合比例的新方法,能够早期监测供体DNA的基因嵌合状态。
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 d
作者
李琳琳
黄一芳
黄运花
潘丽秋
吴祖浩
林发全
Li Linlin;Huang Yifang;Huang Yunhua;Pan Liqiu;Wu Zuhao;Lin Faquan(Department of Clinical Laboratory,First Affiliated Hospital of Guangxi Medical University/Key Laboratory of Clinical Laboratory Medicine of Guangxi Universities,Nanning 530021,China;Guangxi Medical University,Nanning 530021,China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2024年第9期1059-1066,共8页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金(82201913)
广西卫健委自筹课题(Z-A20230482)
广西医科大学大学生创新创业计划(202210598003)。
关键词
Β地中海贫血
造血干细胞移植
第三代测序
嵌合状态
beta-Thalassemia
Hematopoietic stem cell transplantation
Third generation sequencing
Chimerism
