Combining next-generation sequencing and Sanger sequencing for rapid identification of the genetic mutation diagnosis with family tracing of hepatolenticular degeneration
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Combining next-generation sequencing and Sanger sequencing for rapid identification of the genetic mutation diagnosis with family tracing of hepatolenticular degeneration
Journal of Chongqing Medical UniversityVol. 47, Issue 4, Pages: 406-410(2022)
作者机构:
1. 西南医科大学基础医学院生物化学与分子生物学教研室,泸州,646000
2. 中国医学科学院输血研究所分子实验室,成都,610052
3. 四川大学生命科学学院生物资源与生态环境重点实验室,成都,610065
4. 四川大学华西第四医院检验科,成都,610041
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Published:2022,
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Huang Yuan, He Miao, Fan Zhenxin, et al. Combining next-generation sequencing and Sanger sequencing for rapid identification of the genetic mutation diagnosis with family tracing of hepatolenticular degeneration. [J]. Journal of Chongqing Medical University 47(4):406-410(2022)
DOI:
Huang Yuan, He Miao, Fan Zhenxin, et al. Combining next-generation sequencing and Sanger sequencing for rapid identification of the genetic mutation diagnosis with family tracing of hepatolenticular degeneration. [J]. Journal of Chongqing Medical University 47(4):406-410(2022)DOI:
Combining next-generation sequencing and Sanger sequencing for rapid identification of the genetic mutation diagnosis with family tracing of hepatolenticular degeneration
Objective:To investigate an early,accurate and rapid gene detection method and procedure using next-generation sequencing(NGS) full screening and Sanger sequencing based verification for diagnosing a rare genetic disease,hepatolenticular degeneration. Methods:The proband Ⅲ-3 DNA was extracted and the bioinformatic data of high throughout genomic sequences(HTGS) was established to confirm the possible pathogenic sites by using the NGS;then the DNA of all 13 members of the family was extracted,including the proband,in order to design the primer and amplify the target fragment by PCR,then the possible pathogenic sites were verified by using the biopolar generation sequencing. Results:A heterozygous missense mutation(rs121907990;chr13:51937570) was detected on the proband Ⅲ-3’s ATP7B gene exon 8. Three family members,Ⅱ-4,Ⅱ-5 and Ⅲ-4,carried the same heterozygous sense mutation,and their genetic pattern showed mutation located on the same chromosome. The proband’s ATP7B mutation came from the matriarchal origin. Conclusion:The rapid identification method and procedural flow chart designed by our research,which combines the NGS technique and Sanger sequencing based verification technique for the pathogenic site in genealo-gy,has the potential to provide reference for gene diagnosis procedure,and contribute to the timeliness and accuracy of early diagnosis of rare genetic disease.