1. Comparison of Third-Generation Sequencing and Routine Polymerase Chain Reaction in Genetic Analysis of Thalassemia
- Author
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Xu, Zhen, Hu, Lanping, Liu, Yinyin, Peng, Can, Zeng, Guo, Zeng, Li, Yang, Mengyue, Linpeng, Siyuan, Bu, Xiufen, Jiang, Xuanyu, Xie, Tiantian, Chen, Libao, Zhou, Shihao, and He, Jun
- Subjects
Diagnosis ,Usage ,Methods ,Genetic testing -- Methods ,Polymerase chain reaction -- Usage ,DNA sequencing -- Usage ,Thalassemia -- Diagnosis ,Nucleotide sequencing -- Usage ,Genetic screening -- Methods - Abstract
Thalassemia is an inherited disorder characterized by defects in globin synthesis. It is the most widely distributed monogenic autosomal recessive disorder in the world. (1-3) In China, thalassemia is most [...], * Context.--Thalassemia is the most widely distributed monogenic autosomal recessive disorder in the world. Accurate genetic analysis of thalassemia is crucial for thalassemia prevention. Objective.--To compare the clinical utility of a third-generation sequencing-based approach termed comprehensive analysis of thalassemia alleles with routine polymerase chain reaction (PCR) in genetic analysis of thalassemia and explore the molecular spectrum of thalassemia in Hunan Province. Design.--Subjects in Hunan Province were recruited, and hematologic testing was performed. Five hundred four subjects positive on hemoglobin testing were then used as the cohort, and third-generation sequencing and routine PCR were used for genetic analysis. Results.--Of the 504 subjects, 462 (91.67%) had the same results, whereas 42 (8.33%) exhibited discordant results between the 2 methods. Sanger sequencing and PCR testing confirmed the results of third-generation sequencing. In total, third-generation sequencing correctly detected 247 subjects with variants, whereas PCR identified 205, which showed an increase in detection of 20.49%. Moreover, [alpha] triplications were identified in 1.98% (10 of 504) hemoglobin testing-positive subjects in Hunan Province. Seven hemoglobin variants with potential pathogenicity were detected in 9 hemoglobin testing-positive subjects. Conclusions.--Third-generation sequencing is a more comprehensive, reliable, and efficient approach for genetic analysis of thalassemia than PCR, and allowed for a characterization of the thalassemia spectrum in Hunan Province.
- Published
- 2024
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