A multiplex ligationdependent probe amplificationbased nextgeneration sequencing approach for the detection of copy number variations in the human genome.
Mol Med Rep
; 18(6): 5823-5833, 2018 Dec.
Article
in En
| MEDLINE
| ID: mdl-30365071
ABSTRACT
The aim of the present study was to describe a multiplex ligationdependent probe amplification (MLPA)based nextgeneration sequencing (NGS) assay that exhibited a significantly higher efficiency in detecting copy number variations (CNVs) and known singlenucleotide variants, compared with traditional MLPA. MLPA polymerase chain reaction products were used to construct a library with indexed adapters, which was subsequently tested on an NGS platform, and the resulting data were analyzed by a series of analytical software. The reads from each probe reflected genetic variations in the target regions, and fragment differentiation was based on the specific base composition of the sequences, rather than fragment length, which was determined by capillary electrophoresis. The results of this approach were not only consistent with the MLPA results following capillary electrophoresis, but also coincided with the CNV results from the singlenucleotide polymorphism array chip. This method allowed highthroughput screening for the number of fragments and samples by integrating additional indices for detection. Furthermore, this technology precisely and accurately performed largescale detection and quantification of DNA variations, thereby serving as an effective and sensitive method for diagnosing genetic disorders caused by CNVs and known singlenucleotide variations. Notably, MLPANGS circumvents the problems associated with the inaccuracies of NGS in CNV detection due to the use of target sequence capture.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Genome, Human
/
Nucleic Acid Amplification Techniques
/
Genomics
/
DNA Copy Number Variations
/
High-Throughput Nucleotide Sequencing
Type of study:
Diagnostic_studies
Limits:
Child
/
Female
/
Humans
/
Male
Language:
En
Journal:
Mol Med Rep
Year:
2018
Document type:
Article