Detection of single nucleotide polymorphisms in the mannose-binding lectin gene using minor groove binder-DNA probes

• Published in: Journal of immunological methods 2004-04, Vol.287 (1), p.227-230
• Main Authors: Van Hoeyveld, Erna, Houtmeyers, Frans, Massonet, Caroline, Moens, Leen, Van Ranst, Marc, Blanckaert, N., Bossuyt, Xavier
• Format: Article
• Language: English
• Subjects: Allelic discrimination; Biological and medical sciences; DNA Primers; DNA Probes; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genotype; Humans; Mannose-binding lectin; Mannose-Binding Lectin - genetics; MGB probes; Molecular immunology; Molecular Probe Techniques; Point Mutation; Polymerase Chain Reaction - methods; Polymorphism, Single Nucleotide - genetics; Sensitivity and Specificity; Taq Polymerase; Techniques
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2004.01.025

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.