title: Detection of three single nucleotide polymorphisms in the gene encoding mannose-binding lectin in a single pyrosequencing reaction.
abstract: Mannose-binding lectin (MBL) is a key molecule of innate immunity. Binding of MBL to carbohydrates present on pathogens activates the lectin pathway of complement activation, resulting into opsonization and anti-microbial protection. Three frequently occurring single nucleotide polymorphisms (SNPs) are described in the coding region of the MBL2 gene that are associated with abnormal polymerization of the MBL molecule, decreased serum concentrations of high molecular weight MBL, and strongly impaired function. Clinical studies have shown that these MBL SNPs are associated with increased susceptibility to infections, especially in immune-compromised persons, as well as with accelerated progression of chronic diseases. The present study describes a novel method to detect the three major MBL SNPs by pyrosequencing. The close proximity of these SNPs allows their detection in one single pyrosequencing reaction, resulting in clearly distinguishable patterns for each allele combination described until now. This method can be used for the easy and reliable detection of MBL SNPs to identify the basis of functional MBL deficiency in clinical diagnostics and research.
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Postbox 9600, 2300 RC Leiden, The Netherlands. A.Roos@LUMC.NL
authors.forename: Anja
authors.initials: A
authors.lastname: Roos
authors.forename: Patrick
authors.initials: P
authors.lastname: Dieltjes
authors.forename: Rolf H A M
authors.initials: RH
authors.lastname: Vossen
authors.forename: Mohamed R
authors.initials: MR
authors.lastname: Daha
authors.forename: Peter
authors.initials: P
authors.lastname: de Knijff
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005091:Exons; D005838:Genotype; D006801:Humans; D037601:Mannose-Binding Lectin / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: C445619:MBL2 protein, human; D037601:Mannose-Binding Lectin; D004247:DNA