Multiplex Ligation-Dependent Probe Amplification (MLPA)

Multiplex Ligation-dependent Probe Amplification (MLPA) is a multiplex PCR method detecting abnormal copy numbers of specific genes, including small intragenic rearrangements for up to 50 different genomic DNA or RNA sequences. MLPA is a very important technique in the assessment of hereditary diseases including cancers. It is especially indicated whenever large deletions or duplications are described for a gene[5].

What are the advantages of MLPA?

Using MLPA for copy number detection offers many advantages over other techniques: -
• Methods which were primarily developed for detecting point mutations (such as PCR sequencing) generally fail to detect copy numbers changes
• Southern blot analysis will not always detect small deletions and is not ideal as a routine technique
• Although well-characterised deletions and amplifications can be detected by PCR, the exact breakpoint site of most deletions is unknown
• As compared to Fluorescence In Situ Hybridization (FISH), MLPA not only has the advantage of being a multiplex technique, but also one in which very small (50-70 nt) sequences are targeted, enabling MLPA to identify the frequent, single gene aberrations which are too small to be detected by FISH
• MLPA is relatively a low cost and technically uncomplicated method

1. Lovly, C., Berger, M. & Vnencak-Jones, C. (2016). Circulating tumor DNA. My Cancer Genome. Available HERE
2. National Cancer Institute (2017). Liquid biopsy: Using DNA in blood to detect, track, and treat cancer. National Cancer Institute. Available HERE
3. Behjati, S., & Tarpey, P. S. (2013). What is next generation sequencing? Archives of Disease in Childhood. Education and Practice Edition, 98(6), 236–238. LINK
4. The Tayside Centre for Genomic Analysis (2014). Fragment analysis. Available HERE
5. MRC-Holland-MLPA- an introduction. Available HERE