Accurate detection of α-globin gene copy number variants with two reactions using droplet digital PCR

Autor: Xiuqin Bao, Danqing Qin, Jian Ma, Xiangcheng Zhou, Jicheng Wang, Cuize Yao, Liang Zhang, Li Du
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Hematology, Vol 27, Iss 1, Pp 198-203 (2022)
Druh dokumentu: article
ISSN: 1607-8454
16078454
DOI: 10.1080/16078454.2022.2030885
Popis: Background The α-thalassemia is a highly prevalent disease in tropical and subtropical regions, including southern China, and is mainly caused by deletion in α-globin genes (HBA1 and HBA2). The clinical manifestation of α-thalassemia is highly correlated with the copy number of α-globin genes. The decrease in copy number results in α-thalassemia, while duplication or triplication compounded with β-thalassemia may aggravate the clinical manifestation. However, the common methods used to measure the copy number variants can only detect the three common types: –SEA, -α3.7, and -α4.2, and may easily miss the rare deletional type and duplication or triplication cases. Therefore, a new method that allows the detection of different copy number variants in α-globin genes simultaneously and accurately needs to be established. Methods A total of 428 peripheral-blood and fetal chorionic villus or amniotic fluid samples were used in this study. We employed a pair of primers and two probes, one for HBA1 and another for HBA2, to perform droplet digital polymerase chain reaction (ddPCR). Each reaction needed the ddPCR of RPP30 as a reference gene to calculate the copy number. Results We accurately detected the copy number variants in α-globin genes, including the common form α-thalassemia, triplications such as αααanti4.2, and trisomy 16, by performing only two reactions. The accuracy rate for detecting the copy number of α-globin genes was up to 100%. Conclusion In conclusion, ddPCR served as an accurate and rapid method for detecting copy number variations in the clinical screening for α-thalassemia.
Databáze: Directory of Open Access Journals
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