Non-invasive prenatal diagnosis for beta-thalassemia by detecting paternal CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR.
10.3760/cma.j.issn.1003-9406.2018.06.002
- VernacularTitle:用微滴式数字PCR检测母体游离DNA中父源性β-地中海贫血CD41-42突变
- Author:
Yijia ZHANG
1
;
Xiaoqian GONG
;
Yi HE
;
Lichan HUANG
;
Qiang ZHANG
;
Yanhui LIU
;
Jiufeng LI
;
Yajun CHEN
;
Wanjun ZHOU
Author Information
1. Department of Medical Genetics, Southern Medical University, Guangzhou, Guangdong 510515, China. zhouwanjun72@163.com.
- Publication Type:Journal Article
- MeSH:
Cell-Free Nucleic Acids;
DNA;
blood;
Female;
Humans;
Mutation;
Polymerase Chain Reaction;
Pregnancy;
Prenatal Diagnosis;
methods;
beta-Thalassemia;
diagnosis;
genetics
- From:
Chinese Journal of Medical Genetics
2018;35(6):787-790
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish a non-invasive method for beta-thalassemia by detecting parental CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR (ddPCR).
METHODS:Beta-actin gene and beta-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.
RESULTS:The ddPCR method could accurately detect the beta-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=1.0101-X-0.0071 and R=0.9994. The results of 20 maternal plasma cell-free DNA samples were all consistent with those of the follow-up testing.
CONCLUSION:A 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.
