FIG 1.

Schematic diagram of allele-specific PCR (AS-PCR) combined with a lateral flow assay (LFA) system for SNP detection. (A) The workflow of SNP site detection (taking pfmdr1 A256T as an example). Genomic DNA (gDNA) was extracted from a dried filter blood spot (DBS) and used as the template for AS-PCR amplification. After amplification, the PCR products can be directly added to the strips, and they are interpreted according to whether the T line is colored. (B) The SNP detection relies on the wild-type-specific primers and mutant-type-specific primers. Briefly, the wild-type tube includes the wild-type template and primer, and the mutant-type tube includes the mutant-type template and primer. The amplification process will be blocked when the wild-type tube includes the mutant-type template and wild-type primer and when the mutant-type tube includes the wild-type template and mutant-type primer. Both tubes were amplified successfully in the presence of mixed type. WT, Mut, and Mix represent the wild type, mutant type, and mixed type, respectively. (C) Results of pfmdr1 A256T and A551T were analyzed based on agarose gel electrophoresis, DNA sequencing, and signal readout by visual interpretation in LFA. W and M on the sample represent wild-type and mutant-type primers, respectively. C and T represent the control line and test line, respectively.