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C.2678 G>C (GLY893ALA) VARIANT IN COL4A5 GENE IN A PATIENT SUSPECTED TO HAVE ALPORT SYNDROME
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Abstract
Methods: Develop a next-generation sequencing (Illumina) workflow to screen for mutations in the COL4A5, COL4A3, and COL4A4 genes in pediatric patients with suspected Alport nephritis and confirm the findings using Sanger sequencing. Protocol - developing study. Including: DNA extraction from blood samples using the Qiagen (USA) kit, capture hybridized DNA fragments of target genes and construct a sequencing library using the New England Biolabs (USA) kit, sequence using the Illumina (USA) next-generation sequencing system, and confirm the results by Sanger sequencing. Result: A c.2678 G>C (Gly893Ala) mutation in the COL4A5 gene was found, which has not been previously reported in the literature. This mutation is predicted to be the cause of X-linked Alport syndrome based on in silico tools that predict the functional impact of the gene mutation, and it is consistent with the clinical presentation of the patient. Conclusion: The NGS sequencing workflow for detecting mutations in target genes from blood samples of patients with glomerulonephritis has been successfully established. The mutation detected in the COL4A5 gene is a novel mutation in the patient, inherited from the mother.
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Keywords
Col4A5, Alport syndrome, c.2678 G>C (Gly893Ala) mutation
References
2. Boisson, M., et al. (2023). "A wave of deep intronic mutations in X-linked Alport syndrome." Kidney International 104(2): 367-377.
3. Horinouchi, T., et al. (2019). "Determination of the pathogenicity of known COL4A5 intronic variants by in vitro splicing assay." Scientific reports 9(1): 12696.
4. Horinouchi, T., et al. (2018). "Detection of splicing abnormalities and genotype-phenotype correlation in X-linked Alport syndrome." Journal of the American Society of Nephrology 29(8): 2244-2254.
5. Horinouchi, T., et al. (2020). "Pathogenic evaluation of synonymous COL4A5 variants in X‐linked Alport syndrome using a minigene assay." Molecular Genetics & Genomic Medicine 8(8): e1342.
6. Okada, E., et al. (2023). "Aberrant splicing caused by exonic single nucleotide variants positioned 2nd or 3rd to the last nucleotide in the COL4A5 gene." Clinical and Experimental Nephrology 27(3): 218-226.
7. Zhang, R., et al. (2024). "Three exonic variants in the COL4A5 gene alter RNA splicing in a minigene assay." Molecular Genetics & Genomic Medicine 12(2): e2395.