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Date Published: 09/04/2025
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DOI: https://doi.org/10.51298/vmj.v548i1.13304
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Vol. 548 No. 1 (2025)
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Đường Thị Hồng, D., Văn Lộc, V., & Trần Nguyễn Như, U. (2025). APPLYING NEXT GENERATION SEQUENCING (NGS) TO FIND GENE MUTATIONS IN CHILDREN WITH NEPHROTIC SYNDROME: CASE REPORTS. Vietnam Medical Journal, 548(1). https://doi.org/10.51298/vmj.v548i1.13304

APPLYING NEXT GENERATION SEQUENCING (NGS) TO FIND GENE MUTATIONS IN CHILDREN WITH NEPHROTIC SYNDROME: CASE REPORTS

Diệp Đường Thị Hồng, Vũ Văn Lộc, Uyên Trần Nguyễn Như

Main Article Content

Abstract

Objective: To develop a whole exon next-generation sequencing (Illumina - WES) procedure to screen for gene mutations in pediatric patients diagnosed with NS and confirm the results by Sanger sequencing. Methods: Blood samples of 4 pediatric patients diagnosed with NS by a doctor based on clinical manifestations, blood and urine tests. 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 homozygous variant NM_014625.2:c.799G>T (p.Asp267Tyr) in the NPHS2 gene of 2 pediatric patients were detected, it is consistent with the initial diagnosis of corticosteroid-resistant NS by the doctor. In additon, a heterozygous variant c.2858 G>T (Gly953Val) in the COL4A5 gene and heterozygous variant c.3255 G>A (Met1085Ile) in the COL4A3 gene were detected in a pediatric patient, initially diagnosed with focal segmental glomerulosclerosis (FSGS). Futhermore a heterozygous variant c.2630 G>A in the COL4A4 gene of a patient who is being treated and monitored by the doctor for congenital NS. Conclusion: The NGS sequencing workflow for detecting mutations in target genes from blood samples of patients with nephrotic syndrome (NS)  has been successfully established in our laboratory. The sequencing results were analyzed by insilico tools such as Polyphen and SIFT software to predict the function of gene mutations. Alport syndrome can be missed diagnosis.

Article Details

Keywords

Nephrotic syndrome, Alport syndrome, gene COL4A5, COL4A4, COL4A3, gene NPHS2

References

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