A new label-free fluorescent sensor for human. immunodeficiency virus detection based on exonuclease IIIassisted

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1 Electronic Supplementary Material (ESI) for Analyst. This journal is The Royal Society of Chemistry 2016 Supplementary materials for: A new label-free fluorescent sensor for human immunodeficiency virus detection based on exonuclease IIIassisted quadratic recycling amplification and DNAscaffolded silver nanoclusters Wen Yang, Jianniao Tian*, Lijun Wang, Shui Fu, Hongyun Huang, Yanchun Zhao*, Shulin Zhao Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), School of Chemistry and Pharmaceutical Science of Guangxi Normal University, Guilin , China

2 Table S1 Sequences of DNA used in this work Name X-DNA F-DNA Sequences (5 to 3 ) description CCCTTCCGCAGGCACCCCCCCCCCCCCTAACCCTTCCGCAGGCAC CCCCCCCCCCCCTAACAGGTCATGTTATTCCAAATATCTTCT TTAGGGGGGGGGGGGGTGCCTGCGGAAGGGCATTTT B-DNA TTAGGGGGGGGGGGGGTGCCT P-DNA HIV-DNA M1 M3 MN AGGCACCCCCCCCCCCCCTAA AGAAGATATTTGGAATAACATGACCTGGATCCA AGAAGATATTTGGAATAACATCACCTGGATCCA AGAAGATATTTGGAATAACATCTGCTGGATCCA TCTTCTATAAACCTTATTGTACTGGACCTAGGT The underline letters in M1 and M3 are the mismatch based compared with HIV-DNA

3 (F0-F)/F a b c Fig. S1. The performance of amplifying reaction, (a) the quadratic amplifying assay with 1.33µM dsdna, 100 nm HIV-DNA (target), 4 µm P-DNA and 15 U Exo III; (b) single amplifying assay with 100 nm F-DNA (target), 4 µm P-DNA and 15 U Exo III; (c) no amplifying assay with 100 nm B-DNA (target), 4 µm P-DNA and 15 U Exo III. The relative fluorescence intensity was defined as (F 0 -F)/F 0, F 0 represented for the fluorescence intensity of blank sample, F represented for the fluorescence intensity with target. Error bars were calculated from three replicate measurements.

4 (F0-F)/F Concentration of dsdna/μm Fig. S2 Optimization of concentration of dsdna. The relative fluorescence intensity was defined as (F 0 -F)/F 0, F 0 represented for the fluorescence intensity of blank sample, F represented for the fluorescence intensity with HIV-DNA. Error bars were calculated from three replicate measurements. Conditions of reaction: 100 nm HIV-DNA, 4 µm P-DNA and 15 U Exo III.

5 (F0-F)/F Concentrntion of Exo III/ U Fig. S3 Optimization of concentration of Exo III. The relative fluorescence intensity was defined as (F 0 -F)/F 0, F 0 represented for the fluorescence intensity of blank sample, F represented for the fluorescence intensity with HIV-DNA. Error bars were calculated from three replicate measurements. Conditions of reaction: 100 nm HIV-DNA, 4 µm P-DNA and 1.33 µm dsdna.

6 (F0-F)/F Time/ min Fig. S4 Optimization of reaction time of Exo III. The relative fluorescence intensity was defined as (F 0 -F)/F 0, F 0 represented for the fluorescence intensity of blank sample, F represented for the fluorescence intensity with HIV-DNA. Error bars were calculated from three replicate measurements. Conditions of reaction: 100 nm HIV-DNA, 4 µm P-DNA, 1.33 µm dsdna and 15 U Exo III.

7 400 Without HIV With HIV 300 FL Fig. S5 Optimization of systematic buffer. Buffer 1: 20 mm PBS (1mM Mg(Ac) 2, ph 7.0); buffer 2: 20 mm Tris-HAc (70 mm NaAc,10 mm Mg(Ac) 2, 20 mm KAc, ph 7.4 ); buffer 3: 20 mm Tris-HAc (10 mm Mg (Ac) 2, 50 mm KAc, ph 7.9); buffer 4: 10 mm Bis Tris Propane-HCl (10 mm MgCl 2, 1 mm DTT, ph 7.0); buffer 5: 20 mm HEPES-HAc (200 mm NaAc, ph 7.5). FL represented for the fluorescence intensity of the DNA/AgNCs. Error bars were calculated from three replicate measurements. Conditions of reaction: 100 nm HIV-DNA, 4 µm P-DNA, 1.33 µm dsdna and 15U Exo III.

8 Table S2 Comparison of sensitivity by different methods for DNA detection Target Methods labeling Linear range Detection limit Reference DNA Fluorescence No nm Not mentioned 1 HBV Fluorescence No nm 3.0 nm 2 P53 gene Fluorescence Yes nm 50 pm 3 HIV Fluorescence No nm 0.29 nm 4 HBV Polarization Yes nm 0.65 nm 5 DNA Colorimetric No nm 50 pm 6 HBV Fluorescence No nm 0.97 nm 7 DNA Fluorescence No nm Not mentioned 8 DNA Fluorescence No nm 120 pm 9 HIV Fluorescence No 50 pm-5 nm 35 pm This Work Table S3 Recovery of HIV-DNA in spiked human serum samples Samples Added (nm) Found (nm) Recovery (%) RSD (%, n=3)

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