| Salbutamol?SAL?is an adrenergic receptor agonist.Due to environmental residues caused by illegal use,it seriously affects human health,so it is particularly important to establish an ultra-sensitive detection method for it.Some current detection technologies are mainly large-scale instruments,which are difficult to meet on-site needs.A high-sensitivity and specific method to detect salbutamol based on immunomagnetic beads,bridging DNA and two different DNA amplification techniques was established in this paper.First,the complete antigen is modified at the 3 and 5 ends to form adjacent probes,and two adjacent probes are specifically recognized by the antibody coupled to the magnetic beads.Adjacent probes are ligated to form a full-length amplicon,which is used as a template to achieve signal amplification through two DNA amplification techniques.The two DNA signal amplification techniques are real-time fluorescent PCR?qPCR?and strand displacement amplification?SDA?.The result shows that the design of the method has high sensitivity.The linear range of SAL detected by real-time fluorescent quantitative PCR method is 1.0×10-21.0×103 ng·mL-1,and the detection limit is 1.20×10-22 ng·mL-1,the spiked recovery of SAL in tap water samples is between 87.1%and 108.8%.The linear range of SAL detected by strand displacement amplification technique is 1.0×10-31.0×102 pg·mL-1,the detection limit is 2.90 fg·mL-1,and the spiked recovery of SAL in tap water samples is between 93.1%and 103.3%.The two methods have good specificity in their structural functional analogues.This study builds a highly specific and versatile detection platform by designing immunomagnetic beads and bridging DNA to achieve highly sensitive detection of salbutamol under different amplification technologies. |
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