From the Department of Genetics and Eugenics, Changsha Hospital for Maternal & Child Health Care Affiliated to Hunan Normal University, Changsha, China (Xu, Hu, Peng, G. Zeng, L. Zeng, Yang, Linpeng, Bu, Jiang, Zhou, He).
Berry Genomics Corporation, Beijing, China (Liu, Xie, Chen).
Arch Pathol Lab Med. 2024 Mar 1;148(3):336-344. doi: 10.5858/arpa.2022-0299-OA.
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, α 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.
地中海贫血是世界上分布最广泛的单基因常染色体隐性遗传病。地中海贫血的准确基因分析对地中海贫血的预防至关重要。
比较第三代测序技术（全面分析地中海贫血等位基因）与常规聚合酶链反应（PCR）在地中海贫血基因分析中的临床应用，并探讨湖南省地中海贫血的分子谱。
在湖南省招募受试者，并进行血液学检测。然后，将血红蛋白检测呈阳性的 504 名受试者作为研究对象，采用第三代测序和常规 PCR 进行基因分析。
在 504 名受试者中，有 462 名（91.67%）的结果相同，而 42 名（8.33%）的结果在两种方法之间存在差异。Sanger 测序和 PCR 检测证实了第三代测序的结果。第三代测序总共正确检测到 247 名有变异的受试者，而 PCR 只识别到 205 名，检测率提高了 20.49%。此外，在湖南省血红蛋白检测呈阳性的 504 名受试者中，有 1.98%（10 名）检测到α三体。在 9 名血红蛋白检测呈阳性的受试者中，检测到 7 种具有潜在致病性的血红蛋白变异体。
第三代测序是一种比 PCR 更全面、可靠和高效的地中海贫血基因分析方法，能够对湖南省的地中海贫血谱进行特征描述。
