Multiple SNPs Detection Method And Clinial Applications In Phenylketonuria Diagnostic

Single nucleotide polymorphisms(SNPs)are closely related with genetic diseases,whereas current SNPs detection methods such as DNA microarrays including tedious procedures and expensive and sophisticated instruments are no more satisfied with rapid SNPs detection in clinical practice,especially for those multiple SNPs related genetic diseases.In this study,we report a sensitive,low cost and easy-to-use genotyping system formed via combining amplification refractory mutation system(ARMS)polymerase chain reaction with gold magnetic nanoparticles(GMNPs)and lateral flow assay(LFA)noted as ARMS-LFA system,which allow us to use a uniform condition for multiple SNPs detection simultaneously.The genotyping results can be explained by a magnetic reader automatically or be visualized interpretation according to the captured GMNPs probes on the test and control lines of the LFA device.The high sensitivity and specificity of this testing system were found through genotyping seven pathogenic SNPs of phenylalanine hydroxylase(PAH,the etiological factor of phenylketonuria).Furthermore,our ARMS-LFA system was applied to clinical samples for screening 23 families including 69 individuals for the seven pathogenic SNPs of PAH gene.The concordance rate of the genotyping results detected by ARMS-LFA system was up to 97.8% compared with the DNA sequencing results,which make this method a very promising screening method in multiple SNPs caused genetic diseases detection.