Affiliations 

  • 1 Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Center for Neuroscience Services and Research, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
  • 2 Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Department of Pediatrics, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
  • 3 Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
  • 4 Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia
  • 5 Department of Pediatrics, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
  • 6 Hospital Kuala Lumpur, Kuala Lumpur, Malaysia
  • 7 Hospital Sultanah Aminah, Johor Bahru, Malaysia
  • 8 Hospital Raja Perempuan Zainab II, Kota Bharu, Malaysia
  • 9 Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Malaysia
  • 10 Human Genome Center, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Center for Neuroscience Services and Research, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Division of Human Biology, School of Medicine, International Medical University, Kuala Lumpur, Malaysia. Electronic address: teguhhs@usm.my
  • 11 Center for Neuroscience Services and Research, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia; Department of Pediatrics, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
J Mol Diagn, 2017 03;19(2):265-276.
PMID: 28087349 DOI: 10.1016/j.jmoldx.2016.10.009

Abstract

Tuberous sclerosis complex (TSC) is an autosomal dominant neurocutaneous disorder characterized by tumor growth in multiple organs and caused by mutations in either TSC1 or TSC2 genes. Because of their relatively large genomic sizes, absence of hotspots, and common type of mutations, mutation detection in TSC1 and TSC2 genes has been challenging. We devised a combination of multiple ligation-dependent probe amplification (MLPA) and amplicon sequencing (AS) to simplify the detection strategy, yet we come up with reasonably high detection rate. Thirty-four Malaysian patients diagnosed with TSC were referred to Human Genome Center, Universiti Sains Malaysia. We used a combination of MLPA to detect large copy number changes and AS to detect smaller mutations. TSC1 pathogenic or likely pathogenic mutations were found in 6 patients (18%) and TSC2 in 21 patients (62%), whereas 6 patients (18%) show no mutations and 1 patient (2%) showed only TSC2 missense variant with uncertain significance. Six of the mutations are novel. Our detection strategy costs 81% less and require 1 working week less than the conventional strategy. Confirmatory sequencing using Sanger method on a few representative mutations showed agreement with results of the AS. Combination of MLPA and Illumina MiSeq AS provides a simplified strategy and reasonably high detection rate for TSC1/TSC2 mutation, which suggested application of the strategies into clinical molecular diagnostics.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.