Chemiluminescence sensing method based on dsDNA-SYBR Green I light-sensitive catalysis

[0016] Take a certain volume of BRCA1 gene (5'-GAG CAT ACA TAG GGTTTCTCTTGGTTT-3'), 15 μL of 200 nM BRCA1 complementary chain (CS-BRCA1: 5'-AAA CCA AGA GAAACC CTA TGTATG CTC-3') in Add a certain volume of NaH to a multiwell plate with a volume of 300 μL 2 PO 4 After incubating the buffer solution for 60 min, add 2.4 μL of SG dye at a concentration of 250× and incubate for 15 min; then add 15 μL of K at a concentration of 1 mM 4 [Fe(CN) 6 ] solution, placed under a blue LED light (3W) for 30 min. Finally, 100 μL of the sample after illumination was added to 100 μL of 2.0 mM luminol (pH = 11.5). At room temperature, the chemiluminescence intensity was measured with a chemiluminescence instrument, and the concentration of the BRCA1 gene was detected based on the obtained signal intensity.

[0018] Take a certain volume of BRCA2 gene (5'-AAA GGGCTTCTG ATT-3'), 15 μL of BRCA2 complementary strand (CS-BRCA2: 5'-AAT CAG AAG CCC TTT-3') at a concentration of 200 nM in a volume of 300 μL Add a certain volume of NaH to the perforated plate 2 PO 4 After incubating the buffer solution for 60 min, add 2.4 μL of SG dye at a concentration of 250 × and incubate for 15 min; then add 15 μL of K at a concentration of 1 mM 4 [Fe(CN) 6 ] solution, placed under a blue LED light (3W) for 30 min. Finally, 100 μL of the sample after illumination was taken and added with 100 μL of 2.0 mM luminol (pH = 11.5). At room temperature, the chemiluminescence intensity was measured with a chemiluminescence instrument, and the concentration of the BRCA2 gene was detected based on the obtained signal intensity.

[0020] Take a certain volume of p53 gene (5'-TTC CTC TGTGCGCCGGTCTCTCCT-3'), 15 μL of p53 complementary strand (CS-p53: 5'-AGG AGA GACCGGCGCACA GAG GAA-3') at a concentration of 200 nM in a volume of 300 μL Add a certain volume of NaH to the perforated plate 2 PO 4 After incubating the buffer solution for 60 min, add 2.4 μL of SG dye at a concentration of 250 × and incubate for 15 min; then add 15 μL of K at a concentration of 1 mM 4 [Fe(CN) 6 ] solution, placed under a blue LED light (3W) for 30 min. Finally, 100 μL of the sample after illumination was taken and added with 100 μL of 2.0 mM luminol (pH = 11.5). At room temperature, the chemiluminescence intensity was measured with a chemiluminescence instrument, and the concentration of the p53 gene was detected based on the obtained signal intensity.

[0021] The detection limits of BRCA1, BRCA2, and p53 genes detected by Examples 1, 2, and 3 were 3.0 pM, 6.0 pM, and 5.0 pM, respectively. This system can be applied to the...