Snapback Primer Mediated Clamping PCR for Detection of EGFR and KRAS Mutations in NSCLC Patients by High Resolution Melting Analysis
Figure 3
Sensitivity of snapback primer HRM after normal PCR and SPACE-PCR. Mutations accounted for 100%, 1/10, 1/100, 5/1000, 1/1000, 1/104, and 1/105 and wild samples were tested. (a) The sensitivity-test of EGFR E746-A750 del by snapback primer HRM after normal PCR. Lines 1 and 4 are pure wild and mutant, while lines 2 and 4 are 1/100 and 1/10 mutant sample. Here, the 1% detection sensitivity is shown. (b) The sensitivity-test of EGFR E746-A750 del by SPACE-HRM. Lines 1, 2, and 3 stand for wild, 1/105 mutant, and 100% mutant sample, respectively. Now, sensitivity was improved to 1/104. (c) The sensitivity-test of KARS mutation by snapback primer HRM after normal PCR. Lines 1, 2, and 3 stand for 100% mutant, wild, and 1/100 mutant samples; more than 1% of mutant sample can be detected. (d) The sensitivity-test of KARS mutation by SPACE-HRM. Lines 1, 2, and 3 stand for 100% mutant, 1/1000 mutant, and wild samples; more than 1% of mutant sample can be detected. Compared with (a) and (b), SPACE-PCR improves sensitivity of normal snapback primer HRM to 1/1000.
