A SNaPshot Assay for Determination of the Mannose-Binding Lectin Gene Variants and an Algorithm for Calculation of Haplogenotype Combinations

• Main Authors: Jana Mrazkova, Petr Sistek, Jan Lochman, Lydie Izakovicova Holla, Zdenek Danek, Petra Borilova Linhartova
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: Diagnostics
• Subjects: mannose-binding lectin; mannose-binding lectin gene (<i>MBL2)</i>; SNaPshot assay; single nucleotide polymorphism; genotyping; Haplogenotype Calculator
• Online Access: https://www.mdpi.com/2075-4418/11/2/301

Mannose-binding lectin (MBL) deficiency caused by the variability in the <i>MBL2</i> gene is responsible for the susceptibility to and severity of various infectious and autoimmune diseases. A combination of six single nucleotide polymorphisms (SNPs) has a major impact on MBL levels in circulation. The aim of this study is to design and validate a sensitive and economical method for determining <i>MBL2</i> haplogenotypes. The SNaPshot assay is designed and optimized to genotype six SNPs (rs1800451, rs1800450, rs5030737, rs7095891, rs7096206, rs11003125) and is validated by comparing results with Sanger sequencing. Additionally, an algorithm for online calculation of haplogenotype combinations from the determined genotypes is developed. Three hundred and twenty-eight DNA samples from healthy individuals from the Czech population are genotyped. Minor allele frequencies (MAFs) in the Czech population are in accordance with those present in the European population. The SNaPshot assay for <i>MBL2</i> genotyping is a high-throughput, cost-effective technique that can be used in further genetic-association studies or in clinical practice. Moreover, a freely available online application for the calculation of haplogenotypes from SNPs is developed within the scope of this project.