Eczema herpeticum vs dermatitis herpetiformis as a clue of dedicator of cytokinesis 8 deficiency diagnosis: A case report

Figure 3 An overview of multiplex ligation-dependent probe amplification technology for copy number detection. An outline of the MLPA technique: (1) Multiplex ligation-dependent probe amplification (MLPA) probes consist of 2 hemiprobes: a 50 oligonucleotide called the left probe oligonucleotide (LPO) and the 30 oligonucleotide called the right probe oligonucleotide (RPO), which target adjacent sequences (blue) and a polymerase chain reaction (PCR) primer sequence (black). The RPO has a nonhybridizing stuffer sequence (green), which gives the probe a specific length. To begin the reaction, the MLPA probe mix is added to denatured DNA for approximately 16 hours of hybridization; (2) After successful hybridization to their target sequences in the sample DNA, the bound probe oligonucleotides are enzymatically ligated, creating a template for the subsequent PCR reaction; (3) In the PCR reaction, all MLPA probes are amplified by the same PCR primer pair (X and Y), allowing semiquantitative amplification of the probes; (4) Because of the different lengths of the stuffer sequences, the amplification products of different MLPA probes can be separated, identified, and quantified by capillary electrophoresis. This allows the use of up to 50 probes in a single MLPA reaction. Relative amounts of probe amplification products, as compared with a reference DNA sample, reflect the relative copy number of the target sequences. PCR: Polymerase chain reaction; RPO: Right probe oligonucleotide; LPO: Left probe oligonucleotide. Citation: Hömig-Hölzel C, Savola S. Multiplex ligation-dependent probe amplification (MLPA) in tumor diagnostics and prognostics. Diagn Mol Pathol 2012; 21: 189-206. Copyright ©The Authors 2012. Published by Wolters Kluwer Health, Inc.