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BACKGROUND AND PURPOSE: Single nucleotide polymorphism SNP mainly refers to the genomic level by a single nucleotide mutation caused by DNA sequence polymorphisms , genetic variation is the most common mutations . SNP genetic stability, quantity, wide coverage, has been replaced by restriction fragment length polymorphism and microsatellite polymorphism, became the third generation of DNA markers . Usually presents human SNP biallelic polymorphism, can be divided into homozygous normal homozygous and heterozygous genotypes , the development of many diseases associated with a single base mutation is closely related to quickly and accurately detect SNP genotyping of disease , clinical testing and molecular diagnostics of great significance . In this study, β -thalassemia globin gene IVS-2-654 and CD17 which two point mutations as the research object , through the gold nanoparticle technology to improve the detection of asymmetric PCR amplification of specific target sequences obtained , the use of non- labeled nucleic acid probe needle technology, combined ligase fidelity connectivity features to construct a different genotypes were significantly melting point difference between single nucleotide polymorphism SNP detection method. Methods: gold nanoparticles mediated asymmetric PCR target sequence to be detected , to construct a new non- labeled nucleic acid probes , integrated technology and melting point difference ligase expanding technology, through ligation -mediated improved significantly different between genotypes melting point difference , depending on the melting point peak and then quickly and accurately detect and polymorphic SNP genotyping . Results: The suitable concentration of gold nanoparticles improve the efficiency of PCR amplification , by non- labeled nucleic acid probe , the reaction in the connection process, the genotype produced a greater difference in the melting point , to achieve a fast and accurate location mutant genotyping , and the success of the globin gene IVS-2-654 and CD17 which two point mutations in three genotypes for effective genotyping. Conclusion: gold nanoparticle -mediated asymmetric PCR amplification , the PCR yield and efficiency can be improved to obtain a high quality product. The design of a new non- labeled nucleic acid probe , and a melting point by connecting different enzyme technology to expand technology to create a non- fluorescent-labeled SNP detection and classification of the new method. The method is accurate, convenient , fast and low cost , with good fidelity , is expected to genetic disease screening, prevention and genetic diagnosis of some significance
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