Abstract:

Objective To establish polymerase chain reaction（PCR）,allele-specific extension（ASE） and nucleic acid detection strip-based detection for single nucleotide polymorphisms（SNP）. Methods The specific gene sequence containing SNP sites was amplified by PCR. For each SNP site,ASE primers labeled with A were extended. The ASE reaction products can bind to probe labeled with B,and hybridization products containing A and B simultaneously were formed. The products can be detected by disposable amplicon cross-contamination proof device containing a nucleic acid detection strip,and the detection of SNP genotypes can be accomplished. Results Ten-fold serial dilutions of quantified human genomic DNA were used to determine the sensitivity of PCR-ASE（88 ng/reaction）. The ability of duplex PCR-ASE with the products can be detected by a single nucleic acid detection strip. A total of 19 samples representing 5 common SNP were detected by PCR-ASE,and the results had the consistency of 100% with DNA sequencing. Conclusions PCR-ASE is simple and accurate detection for SNP.

Key words: Allele-specific extension, Single nucleotide polymorphism, Nucleic acid detection strip-based detection