Abstract
Background: Multiplex ligation-dependent probe amplification (MLPA) is a novel method that uses multiple pairs of oligonucleotide probes to hybridize targeted genomic DNA regions; products from ligation reaction and polymerase chain reaction amplification are then analyzed by capillary electrophoresis. MLPA has been broadly used to detect copy number variation from single nucleotide substitutions to several megabase nucleotide deletions/duplications to chromosomal aneuploidy, methylation pattern analysis for genetic imprinting disorders and malignant tumors, and gene expression profiles in various genetic conditions. Objective: The technical aspects of the MLPA assay are reviewed, its applications in molecular diagnosis are discussed, and information is presented about quality assurance, quality control and future possibilities. Methods: More than 200 MLPA-related publications have been reviewed. Conclusion: MLPA offers significant advantages and is an attractive alternative to many current molecular and cytogenetic diagnostic techniques for broad applications. For its robustness and potential high throughput, it is anticipated that MLPA technology will be more widely used in the molecular diagnostic laboratory as a routine test for many genetic disorders.
Acknowledgements
The authors thank C-S Jonathan Liu (SoftGenetics) for discussions on data analysis, K Sumner for preparing whole genome amplified samples, J McCowen-Rose for critical editing of this manuscript and MRC-Holland for technical assistance of MLPA.