Using KASP technique to screen LRRK2 G2019S mutation in a large Tunisian cohort

Abstract

BackgroundIn North African populations, G2019S mutation in LRRK2 gene, encoding for the leucine-rich repeat kinase 2, is the most prevalent mutation linked to familial and sporadic Parkinson’s disease (PD). Early detection of G2019S by fast genetic testing is very important to guide PD’s diagnosis and support patients and their family caregivers for better management of their life according to disease’s evolution.MethodsIn our study, a genetic PD’s diagnosis tool was developed for large scale genotyping using Kompetitive Allele Specific PCR (KASP) technology. We investigated G2019S’s frequency in 250 Tunisian PD patients and 218 controls.ResultsWe found that 33.6% of patients and 1.3% of controls were carriers. Demographic characteristics of patients with G2019S had no differences compared with non-carrier patients. Thereby, we could emphasize the implication of G2019S in PD without any distinctive demographic factors in the studied cohort. Sixty patients out of 250 were genotyped using Taqman assay and Sanger sequencing. The genotyping results were found to be concordant with KASP assay.ConclusionsThe G2019S mutation frequency in our cohort was similar to that reported in previous studies. Comparing to Taqman assay and Sanger sequencing, KASP was shown to be a reliable, time and cost effective genotyping assay for routine G2019S screening in genetic testing laboratories.

Kompetitive Allele Specific PCR (KASP™) genotyping of 48 polymorphisms at different caprine loci in French Alpine and Saanen goat breeds and their association with milk composition

“…Ours results are in accordance with previously reported KASP assay success rate ( Semagn et al, 2014 ). Furthermore, according to previous studies the KASP genotyping system has low-labour and economic advantages over other genotyping assays ( Landoulsi et al, 2017 ).…”

“…The Kompetitive Allele Specific PCR genotyping assay has been successfully used in order to genotype different organisms ( He, Holme & Anthony, 2014 ; Semagn et al, 2014 ; Landoulsi et al, 2017 ). In our work KASP technology was used in Saanen and Alpine goats reared in Romania in order to investigate the polymorphism of 48 SNPs of 40 genes that were previously reported to be associated with the production traits or that potentially contributed to important metabolic conditions.…”

Characterization of ADME genes variation in Roma and 20 populations worldwide

“…The KASP genotyping assay is a form of competitive allele-specific PCR combined with homogeneous fluorescent SNP genotyping system, which determines the alleles at a specific locus within genomic DNA [ 53 ]. This technology has been widely used on plant species, while recently it has been successfully applied to human samples too [ 54 , 55 ]. From the list of evidence-proved genetic biomarkers associated with metabolism of drugs, which is available at www.pharmaadme.org , 137 single nucleotide polymorphisms (SNPs) were selected for genotyping using the KASP and 127 of them were genotyped successfully.…”

Comparison of three PCR-based assays for SNP genotyping in plants

“…The TaqMan and the KASP assays have been widely used for genotyping due to their high-throughput, low cost, sensitivity and tolerance of variation in the quality and quantity of input DNA. In addition, the TaqMan and KASP assays are suitable for use on a variety of real-time PCR instruments [ 10 ]. However, these platforms still suffer from imperfect sensitivity and allele discrimination, along with less expensive costs of implementation.…”