Original Article
Non-invasive prenatal diagnosis for β-thalassemia by detecting paternal CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR
Yijia Zhang, Xiaoqian Gong, Yi He, Lichan Huang, Qiang Zhang, Yanhui Liu, Jiufeng Li, Yajun Chen, Wanjun Zhou
Published 2018-12-10
Cite as Chin J Med Genet, 2018, 35(6): 787-790. DOI: 10.3760/cma.j.issn.1003-9406.2018.06.002
Abstract
ObjectiveTo establish a non-invasive method for β-thalassemia by detecting parental CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR (ddPCR).
Methodsβ-actin gene and β-thalassemia gene CD41-42 mutation were respectively set as the reference and target sequences. A novel method was established based on Bio-Rad ddPCR technique with specific primers and TaqMan probes for the two genes. The accuracy, sensitivity and detective linearity range of the developed method were evaluated by detection of the target gene gradient concentration samples. The applicability was also evaluated by testing 20 maternal plasma samples.
ResultsThe ddPCR method could accurately detect the β-thalassemia CD41-42 mutation in cell-free DNA derived from maternal plasma. Within the target sequence concentration ratio of 5.00%-0.50%, the relative errors were all <0.05, the linear regression equation was Y=1.0101X -0.0071 and R2=0.9994. The results of 20 maternal plasma cell-free DNA samples were all consistent with those of the follow-up testing.
ConclusionA ddPCR method for detecting parental CD41-42 mutation in cell-free DNA from maternal plasma was developed. The method is simple, rapid, accurate, and can be applied for non-invasive prenatal diagnosis for couples simultaneously carrying the CD41-42 mutation.
Key words:
Droplet digital PCR; Thalassemia; Prenatal diagnosis; Gene mutation
Contributor Information
Yijia Zhang
Department of Medical Genetics, Southern Medical University, Guangzhou, Guangdong 510515, China
Xiaoqian Gong
Shaoguan Maternal and Child Health Care Hospital, Shaoguan, Guangdong 512026, China
Yi He
Dongguan Maternal and Child Health Care Hospital, Dongguan, Guangdong 523000, China
Lichan Huang
Shaoguan Maternal and Child Health Care Hospital, Shaoguan, Guangdong 512026, China
Qiang Zhang
Department of Medical Genetics, Southern Medical University, Guangzhou, Guangdong 510515, China
Yanhui Liu
Dongguan Maternal and Child Health Care Hospital, Dongguan, Guangdong 523000, China
Jiufeng Li
Shaoguan Maternal and Child Health Care Hospital, Shaoguan, Guangdong 512026, China
Yajun Chen
Shaoguan Maternal and Child Health Care Hospital, Shaoguan, Guangdong 512026, China
Wanjun Zhou
Department of Medical Genetics, Southern Medical University, Guangzhou, Guangdong 510515, China
