Genotyping Of Single Nucleotide Polymorphisms (Snps) Of Ctla-4 And Foxp3 Genes In Lymphatic Filariasis And Schistosomiasis
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Date
2011-02
Authors
Md Idris, Zulkarnain
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Lymphatic filariasis (LF) is a chronic parasitic disease caused by thread-like worms of Brugia and Wuchereria species that live in the human lymphatic system. It is a major cause of permanent disability and disfigurement in the endemic areas. Schistosomiasis is another chronic condition caused by schistosoma species which remains as a prevalent parasitic disease worldwide; it causes damage to internal organs and impair growth in children. Susceptibility to infections and disease severities are associated with a wide range of human genetic variations. However such association has not been well investigated especially with regard to CTLA-4 and FoxP3 genes expressed by Tregs, for their functional roles and contributions to the risk of chronic helminth infections. SNP genotyping approach was used in this study to identify potential SNPs in LF caused by Brugia malayi and schistosomiasis caused by Schistosoma haematobium. Briefly the methodology involved extraction and quantification of DNA samples from both parasitic diseases and healthy controls from the endemic areas. Subsequently, using five SNPs in CTLA-4 gene and one SNP in FoxP3 gene, samples for positive controls were selected by performing small scale real-time PCR genotyping on ten samples followed by confirmation by DNA sequencing. This was followed by large scale genotyping of all samples. Finally, analyses of the genotypes were performed using several statistical methods. Genotype confirmation by direct sequencing of three positive controls namely 6944W, 6725H and 20M was found to be identical with real-time PCR. Out of six positive control samples, four samples (6725H, 6944W, 8209H and 20M) with insufficient DNA material were successfully cloned.
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Keywords
Single Nucleotide Polymorphisms (Snps) , Ctla-4 And Foxp3 Genes