Two-color multiplex ligation-dependent probe amplification: detecting genomic rearrangements in hereditary multiple exostoses

Human Mutation
Stefan J WhiteJ T den Dunnen

Abstract

Genomic deletions and duplications play an important role in the etiology of human disease. Versatile tests are required to detect these rearrangements, both in research and diagnostic settings. Multiplex ligation-dependent probe amplification (MLPA) is such a technique, allowing the rapid and precise quantification of up to 40 sequences within a nucleic acid sample using a one-tube assay. Current MLPA probe design, however, involves time-consuming and costly steps for probe generation. To bypass these limitations we set out to use chemically synthesized oligonucleotide probes only. The inherent limitations of this approach are related to oligonucleotide length, and thus the number of probes that can be combined in one assay is also limited. This problem was tackled by designing a two-color assay, combining two sets of probes, each amplified by primers labeled with a different fluorophore. In this way we successfully combined 28 probes in a single reaction. The assay designed was used to screen for the presence of deletions and duplications in patients with hereditary multiple exostoses (HME). Screening 18 patients without detectable point mutations in the EXT1 and EXT2 genes revealed five cases with deletions of one or more ex...Continue Reading

Associated Clinical Trials

Oct 21, 2019·Luca Sangiorgi

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Citations

Jun 29, 2007·Human Genetics·Cornelis L HarteveldPiero C Giordano
Sep 24, 2005·European Journal of Medical Genetics·Marijke BautersGuy Froyen
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Mar 16, 2007·Journal of Medical Genetics·Junko KannoShigeo Kure
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