Multiplex Ligation-dependent Probe Amplification (MLPA) et sondes « à façon » entièrement synthétiques. Guide pratique, recommandations et expérience au CHU de Grenoble

Multiplex Ligation-dependent Probe Amplification (MLPA) is a powerful technique for gene dosage quantification. This technique is based on the ligation followed by PCR amplification of two adjacently hybridising oligonucleotides. Its reliability and effectiveness have been proven and is now well established. Besides the commercial kits are distributed by the patentee, MRC-Holland™, it makes it possible to utilise custom-designed MLPA probes for a targeted use of this technique. This strategy has opened the field of MLPA for various applications ranging from diagnostic of rarest pathologies to confirmation of a CGH-detected abnormalities. However, it may be difficult to develop without a solid experience. Here, we detail the guidelines and recommendations for designing custom MLPA probes and illustrate their use in our laboratory.