ANALYSIS OF GENE COPY NUMBER VARIATIONS IN PATIENTS WITH CARDIAC SEPTAL DEFECTS USING MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION: CNVs analysis in CSDs

YASHVANTHI BORKAR; KRISHNANANDA NAYAK; RANJAN SHETTY K; RAJASEKHAR MOKA

doi:10.22159/ajpcr.2020.v13i1.36189

Authors

YASHVANTHI BORKAR Department of Cell and Molecular Biology, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, India.
KRISHNANANDA NAYAK Department of Cardiovascular Technology, Manipal College of Allied Health Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, India.
RANJAN SHETTY K Department of Cardiology, Kasturba Hospital, Manipal Academy of Higher Education, Manipal, Karnataka, India.
RAJASEKHAR MOKA Department of Cell and Molecular Biology, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, India

DOI:

https://doi.org/10.22159/ajpcr.2020.v13i1.36189

Keywords:

Copy number variations, Atrial septal defects, Ventricular septal defects, Multiplex ligation-dependent probe amplification, Heterozygous deletions, Heterozygous duplications

Abstract

Objective: Cardiac septal defects (CSDs), the most common human congenital heart malformations are complex and heterogeneous. Progress in molecular biology has helped to identify many genes responsible for cardiac morphogenesis. However, etiologic factors in familial as well as isolated syndromes are being identified; the root genetic cause still needs to be resolved and its mechanism is yet to be revealed. The objective of this study is to identify DNA copy number variations (CNVs) and their possible association with septal defects.

Methods: Multiplex ligation-dependent probe amplification (MLPA) was used to detect DNA copy number in non-syndromic CSDs using the P311-A1 Kit consisting of probes for the key genes, namely, NKX2-5 (NK2 transcription factor related, locus 5), GATA4 (GATA binding protein 4), TBX5 (T-box transcription factor), bone morphogenetic protein 4, and CRELD1 (cysteine rich with EGF-like domains 1).

Results: We studied 124 clinically diagnosed CSD subjects, of which 111 (89.5%) had atrial septal defects and 13 (10.5%) had ventricular septal defects. MLPA assay was carried out in all these patients after a thorough clinical and cytogenetic screening. CNVs were identified in 16 (12.9%) cases, of which heterozygous deletions and heterozygous duplications were detected (8 patients each) with apparent phenotypes.

Conclusion: MLPA could be a useful assay for the detection of CNVs and to be adopted as the first line of screening in patients with congenital heart diseases.

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