Haitao Wu1, Dongjia Chen2, Qiang Zhao1, Xiaoting Shen2, Yongbin Liao1, Ping Li1, Philip C N Chiu3,4, Canquan Zhou5. 1. Reproductive Medicine Center, Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-Sen University, Jiangmen, Guangdong, China. 2. Reproductive Medicine Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China. 3. Department of Obstetrics and Gynaecology, Queen Mary Hospital, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China. pchiucn@hku.hk. 4. Shenzhen Key Laboratory of Fertility Regulation, Department of Obstetrics and Gynecology, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China. pchiucn@hku.hk. 5. Reproductive Medicine Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China. zhoucanquan@mail.sysu.edu.cn.
Abstract
PURPOSE: This study aimed to evaluate the value of long-read sequencing for preimplantation haplotype linkage analysis. METHODS: The genetic material of the three β-thalassemia mutation carrier couples was sequenced using single-molecule real-time sequencing in the 7.7-kb region of the HBB gene and a 7.4-kb region that partially overlapped with it to detect the presence of 17 common HBB gene mutations in the Chinese population and the haplotypes formed by the continuous array of single-nucleotide polymorphisms linked to these mutations. By using the same method to analyze multiple displacement amplification products of embryos from three families and comparing the results with those of the parents, it could be revealed whether the embryos carry disease-causing mutations without the need for a proband. RESULTS: The HBB gene mutations of the three couples were accurately detected, and the haplotype linked to the pathogenic site was successfully obtained without the need for a proband. A total of 68.75% (22/32) of embryos from the three families successfully underwent haplotype linkage analysis, and the results were consistent with the results of NGS-based mutation site detection. CONCLUSION: This study supports long-read sequencing as a potential tool for preimplantation haplotype linkage analysis.
PURPOSE: This study aimed to evaluate the value of long-read sequencing for preimplantation haplotype linkage analysis. METHODS: The genetic material of the three β-thalassemia mutation carrier couples was sequenced using single-molecule real-time sequencing in the 7.7-kb region of the HBB gene and a 7.4-kb region that partially overlapped with it to detect the presence of 17 common HBB gene mutations in the Chinese population and the haplotypes formed by the continuous array of single-nucleotide polymorphisms linked to these mutations. By using the same method to analyze multiple displacement amplification products of embryos from three families and comparing the results with those of the parents, it could be revealed whether the embryos carry disease-causing mutations without the need for a proband. RESULTS: The HBB gene mutations of the three couples were accurately detected, and the haplotype linked to the pathogenic site was successfully obtained without the need for a proband. A total of 68.75% (22/32) of embryos from the three families successfully underwent haplotype linkage analysis, and the results were consistent with the results of NGS-based mutation site detection. CONCLUSION: This study supports long-read sequencing as a potential tool for preimplantation haplotype linkage analysis.
Authors: Robert Pinard; Alex de Winter; Gary J Sarkis; Mark B Gerstein; Karrie R Tartaro; Ramona N Plant; Michael Egholm; Jonathan M Rothberg; John H Leamon Journal: BMC Genomics Date: 2006-08-23 Impact factor: 3.969
Authors: Aaron M Wenger; Paul Peluso; William J Rowell; Pi-Chuan Chang; Richard J Hall; Gregory T Concepcion; Jana Ebler; Arkarachai Fungtammasan; Alexey Kolesnikov; Nathan D Olson; Armin Töpfer; Michael Alonge; Medhat Mahmoud; Yufeng Qian; Chen-Shan Chin; Adam M Phillippy; Michael C Schatz; Gene Myers; Mark A DePristo; Jue Ruan; Tobias Marschall; Fritz J Sedlazeck; Justin M Zook; Heng Li; Sergey Koren; Andrew Carroll; David R Rank; Michael W Hunkapiller Journal: Nat Biotechnol Date: 2019-08-12 Impact factor: 54.908