| Autor: |
Ismail, Nur Farrah Dila, Rani, Abdul Qawee, Nik Abdul Malik, Nik Mohd Ariff, Boon Hock, Chia, Mohd Azlan, Siti Nabilahuda, Abdul Razak, Salmi, Keng, Wee Teik, Ngu, Lock Hock, Silawati, Abdul Rashid, Yahya, Nor Azni, Mohd. Yusoff, Narazah, Sasongko, Teguh Haryo, Zabidi-Hussin, Z.A.M.H. |
| Zdroj: |
The Journal of Molecular Diagnostics; March 2017, Vol. 19 Issue: 2 p265-276, 12p |
| Abstrakt: |
Tuberous sclerosis complex (TSC) is an autosomal dominant neurocutaneous disorder characterized by tumor growth in multiple organs and caused by mutations in either TSC1or TSC2genes. Because of their relatively large genomic sizes, absence of hotspots, and common type of mutations, mutation detection in TSC1and TSC2genes 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. TSC1pathogenic or likely pathogenic mutations were found in 6 patients (18%) and TSC2in 21 patients (62%), whereas 6 patients (18%) show no mutations and 1 patient (2%) showed only TSC2missense 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/TSC2mutation, which suggested application of the strategies into clinical molecular diagnostics. |
| Databáze: |
Supplemental Index |
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