Min Chen1,2,3,4, Chao Chen5, Xiaoyan Huang6, Jun Sun5, Lu Jiang5, Yingting Li1, Yaping Zhu5, Changgeng Tian6, Yufan Li1,2,3,4, Zhe Lu5, Yaoshen Wang5, Fanwei Zeng6, Yun Yang7, Xiwei Song6, Zhiyu Peng6, Chenghong Yin8, Dunjin Chen1,2,3,4,9. 1. Department of Fetal Medicine and Prenatal Diagnosis, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China. 2. Obstetrics and Gynecology Institute of Guangzhou, Guangzhou, China. 3. The Medical Centre for Critical Pregnant Women in Guangzhou, Guangzhou, China. 4. Key Laboratory for Major Obstetric Diseases of Guangdong Province, Guangzhou, China. 5. Tianjin Medical Laboratory, BGI-Tianjin, BGI-Shenzhen, Tianjin, China. 6. BGI Genomics, BGI-Shenzhen, Shenzhen, China. 7. Wuhan BGI Clinical Laboratory Co., Ltd, BGI-Wuhan, BGI-Shenzhen, Wuhan, China. 8. Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China. 9. Key Laboratory for Reproduction and Genetics of Guangdong Higher Education Institutes, Guangzhou, China.
Abstract
OBJECTIVE: We aimed to investigate the validity of noninvasive prenatal diagnosis (NIPD) based on direct haplotype phasing without the proband or other family members and its feasibility for clinical application in the case of Duchenne muscular dystrophy (DMD). METHODS: Thirteen singleton-pregnancy families affected by DMD were recruited. The pathogenic variants in the pregnant females have been identified by multiplex ligation-dependent probe amplification (MLPA). We resolved maternal haplotypes for each family by performing targeted linked-read sequencing of their high molecular weight DNA, respectively. Then, we integrated the maternal haplotypes and the targeted sequencing results of maternal plasma DNA to infer the fetal haplotype and the DMD gene variant status. The fetal genotypes were further validated by using chorionic villus sampling. RESULTS: The method of directly resolving maternal haplotype through targeted linked-read sequencing was smoothly performed in 12 participated families, but one failed (F11). The predicted variant status of 12 fetuses was correct, which had been confirmed by invasive prenatal diagnosis. CONCLUSION: Direct haplotyping of NIPD based on linked-read sequencing for DMD is accurate.
OBJECTIVE: We aimed to investigate the validity of noninvasive prenatal diagnosis (NIPD) based on direct haplotype phasing without the proband or other family members and its feasibility for clinical application in the case of Duchenne muscular dystrophy (DMD). METHODS: Thirteen singleton-pregnancy families affected by DMD were recruited. The pathogenic variants in the pregnant females have been identified by multiplex ligation-dependent probe amplification (MLPA). We resolved maternal haplotypes for each family by performing targeted linked-read sequencing of their high molecular weight DNA, respectively. Then, we integrated the maternal haplotypes and the targeted sequencing results of maternal plasma DNA to infer the fetal haplotype and the DMD gene variant status. The fetal genotypes were further validated by using chorionic villus sampling. RESULTS: The method of directly resolving maternal haplotype through targeted linked-read sequencing was smoothly performed in 12 participated families, but one failed (F11). The predicted variant status of 12 fetuses was correct, which had been confirmed by invasive prenatal diagnosis. CONCLUSION: Direct haplotyping of NIPD based on linked-read sequencing for DMD is accurate.
Authors: C Liautard-Haag; G Durif; C VanGoethem; D Baux; A Louis; L Cayrefourcq; M Lamairia; M Willems; C Zordan; V Dorian; C Rooryck; C Goizet; A Chaussenot; L Monteil; P Calvas; C Miry; R Favre; E Le Boette; M Fradin; A F Roux; M Cossée; M Koenig; C Alix-Panabière; C Guissart; M C Vincent Journal: Sci Rep Date: 2022-07-06 Impact factor: 4.996