Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (12): 81-89.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0698
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HOU Wei-chen1(), YE Ke1, LI Jie2, ZHANG Yang-zi3, XU Wen-tao3, ZHU Long-jiao3, LI Xiang-yang1()
Received:
2023-07-19
Online:
2023-12-26
Published:
2024-01-11
Contact:
LI Xiang-yang
E-mail:houweichen007@163.com;lxy2002cn@163.com
HOU Wei-chen, YE Ke, LI Jie, ZHANG Yang-zi, XU Wen-tao, ZHU Long-jiao, LI Xiang-yang. Detection of Escherichia coli O157: H7 Based on Antibody Aptamer Sandwich Biosensor[J]. Biotechnology Bulletin, 2023, 39(12): 81-89.
名称 Name | 序列 Sequence(5'-3') |
---|---|
S1 | CAG GTC CAT CGA GTG GTA GGA TGG TCG TGG TGA GGT GCG TGT ATG GGT GGT GGA TGA GTG TGT GGC TCG CAC TGC TCC TGA ACG TAC |
S1-1 | CAG GTC CAT CGA GTG GTA GGA TGG TCG TGG TGA GGT GCG TGT A TCG CAC TGC TCC TGA ACG TAC |
S1-2 | CAG GTC CAT CGA GTG GTA GGA TGG GTG GTG GAT GAG TGT GTG GC TCG CAC TGC TCC TGA ACG TAC |
S1-3 | CAG GTC CAT CGA GTG GTA GGA GAG GTG CGT GTA TGG GTG GT TCG CAC TGC TCC TGA ACG TAC |
S1-3-1 | CAG GTC CAT CGA GTG GTA GGA GAG GTG CGT GTA TCG CAC TGC TCC TGA ACG TAC |
FP | CAG GTC CAT CGA GTG GTA GGA |
RP | GTA CGT TCA GGA GCA GTG CGA |
Table 1 Nucleic acid sequences used in the experiment
名称 Name | 序列 Sequence(5'-3') |
---|---|
S1 | CAG GTC CAT CGA GTG GTA GGA TGG TCG TGG TGA GGT GCG TGT ATG GGT GGT GGA TGA GTG TGT GGC TCG CAC TGC TCC TGA ACG TAC |
S1-1 | CAG GTC CAT CGA GTG GTA GGA TGG TCG TGG TGA GGT GCG TGT A TCG CAC TGC TCC TGA ACG TAC |
S1-2 | CAG GTC CAT CGA GTG GTA GGA TGG GTG GTG GAT GAG TGT GTG GC TCG CAC TGC TCC TGA ACG TAC |
S1-3 | CAG GTC CAT CGA GTG GTA GGA GAG GTG CGT GTA TGG GTG GT TCG CAC TGC TCC TGA ACG TAC |
S1-3-1 | CAG GTC CAT CGA GTG GTA GGA GAG GTG CGT GTA TCG CAC TGC TCC TGA ACG TAC |
FP | CAG GTC CAT CGA GTG GTA GGA |
RP | GTA CGT TCA GGA GCA GTG CGA |
Fig. 2 Feasibility analysis of adaptor clipping A:Amplification curve of original bacterial solution adaptor and gradient dilution.B:Change trend of original bacterial solution adaptor and gradient dilution
Fig. 3 Comparison of tailored adaptor and original adaptor A: S1 and S1-1 amplification curves; B: S1 and S1-2 amplification curves; C: S1 and S1-3 amplification curves; D: S1 and S1-3-1 amplification curves
Fig. 5 Feasibility analysis for biosensor A: Amplification curves of three complexes of IMB target adaptor, IMB adaptor, and IMB target. B: Amplification curves after four times ten fold gradient dilution of bacterial solution
Fig. 6 Condition optimization results A: Optimization of the type of adapted body scissor binding buffer; B: optimization of the pH of adapted body scissor buffer; C: optimization of adapted body scissor incubation temperature; D: optimization of sensor IMB target incubation time
样品 Sample | 传统培养法Traditional cultivation method /(CFU·mL-1) | 检测量Detection quantity/ (CFU·mL-1) | 回收率 Rate of recovery/% | RSD/% |
---|---|---|---|---|
1 | 3×103 | 2.89×103 | 96.3 | 2.15 |
2 | 3×104 | 3.11×104 | 103 | 2.07 |
3 | 3×105 | 3.26×105 | 108 | 1.35 |
Table 2 Actual sample testing
样品 Sample | 传统培养法Traditional cultivation method /(CFU·mL-1) | 检测量Detection quantity/ (CFU·mL-1) | 回收率 Rate of recovery/% | RSD/% |
---|---|---|---|---|
1 | 3×103 | 2.89×103 | 96.3 | 2.15 |
2 | 3×104 | 3.11×104 | 103 | 2.07 |
3 | 3×105 | 3.26×105 | 108 | 1.35 |
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