Multiplex Ligation-dependent Probe Amplification (MLPA) is a method to determine the copy number of up to 60 genomic DNA sequences in a single multiplex PCR based reaction. MLPA probes consist of two oligonucleotides that can hybridize next to each other on a certain DNA sequence of interest, where they are ligated. All ligated probes are subsequently amplified by PCR using a single set of primers. Each amplified MLPA probe has a unique length and can be visualized and quantified by capillary electrophoresis. As the primers are almost 100% consumed in the PCR reaction, the quantity of each PCR amplicon is proportional to the number of copies of each probe target sequence in the DNA sample. A trisomy 21 can therefore be detected by an approximately 50% increased signal of each chromosome 21 specific probe relative to reference samples. MLPA with the P095 Aneuploidy probemix for chromosomes 13, 18, 21, X and Y has been used as a rapid detection method on large numbers of samples from uncultured amniotic fluid or from chorionic villi. As compared to FISH and karyotyping, MLPA is more rapid, has a higher throughput, and is less expensive. MLPA however cannot detect low grade mosaicism, female triploidies, and copy number neutral chromosome abnormalities such as inversions and translocations.

Amniotic fluid, Aneuploidy, Chorionic villi, Gene dosage, Multiplex ligation-dependent probe amplification (MLPA), Multiplex polymerase chain reaction (PCR), Trisomy,
Methods in Molecular Biology
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Department of Clinical Genetics

Schouten, J.P, van Vught, P.W.J, & Galjaard, R-J.H. (2019). Multiplex ligation-dependent probe amplification (MLPA) for prenatal diagnosis of common aneuploidies. In Methods in Molecular Biology. doi:10.1007/978-1-4939-8889-1_11