Allele-Specific Real-Time Polymerase Chain Reaction as a Tool for Urate Transporter 1 Mutation Detection

Abstract

Allele-specific polymerase chain reaction (ASPCR) method has long been applied for the detection of nucleotide variations and genotyping, which are detected by the presence or absence of DNA amplification PCR products. Recently, Real-Time PCR genotyping has fast developed and offered a rapid method of detecting mutations without the need of gel electrophoresis as with ASPCR. Here, we describe an easy and rapid touchdown real-time PCR method for the detection of nucleotide variations. Using our method we successfully detect two main mutations in human urate transporter 1 (SLC22A12), W258X and R90H, and validate the results. The method can potentially be applied to genotype of various other nucleotide variations.

Application of nested-qPCR-High resolution melting (HRM) technology on Strongyloides stercoralis isolates from Iran

“…Although, this method cannot be a proper substitute for sequencing [18], but it is able to scan even single nucleotide polymorphisms (SNPs) in DNA [21]. It has also some advantages such as rapid screening and detection of closely related species in a laboratory [15]. In the present study, the nested-qPCR-HRM assay based on cox1 gene ampli cation was performed on some human isolates of S. stercoralis from four endemic provinces of Iran.…”

“…High resolution melting (HRM) analysis is a multipurpose technology, which attracts a range of new users due to its ease of use, simplicity, exibility, low cost, and high sensitivity and speci city [15]. Recently HRM technology has been applied for screening of the patients infected with various helminth infection such as strongyloidosis [16], Trichostrongylus spp.…”