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  • Title: Detection of single nucleotide polymorphisms in the mannose-binding lectin gene using minor groove binder-DNA probes.
    Author: Van Hoeyveld E, Houtmeyers F, Massonet C, Moens L, Van Ranst M, Blanckaert N, Bossuyt X.
    Journal: J Immunol Methods; 2004 Apr; 287(1-2):227-30. PubMed ID: 15099770.
    Abstract:
    Structural point mutations in exon 1 at codons 52, 54 and 57 and a promotor polymorphism at -221 bp of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to various infectious diseases. We developed a genotyping method based on the 5' nuclease (TaqMan) assay in combination with the use of minor-groove-binder (MGB) probes in screening for these mutations/polymorphisms. In contrast to conventional probes, MGB probes have a short length and can be used for detection of mutations that are in close proximity to each other, as is the case for the structural mutations in exon 1 of the MBL gene. Results obtained with the 5' nuclease assay using MGB probes were identical with results obtained with classical techniques such as restriction fragment length polymorphism, allele-specific PCR, and sequencing. In conclusion, the 5' nuclease assay using MGB probes is useful for large-scale screening of point mutations/polymorphisms, even when they are in close proximity.
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