The invention discloses an
aptamer-based
oxytetracycline fluorescent quantitative PCR (
Polymerase Chain Reaction) detection kit as well as a preparation method and a detection method thereof, and relates to the technical field of analysis and detection.The detection method comprises the following steps: mixing and incubating a sample to be detected, an
oxytetracycline specific
aptamer, a complementary chain and a
magnetic bead curing capture probe; magnetically separating out the mixed and incubated magnetic beads, curing and capturing the probe, and washing to obtain a magnetic compound; adding PCR reaction liquid containing the primer pair and a
TaqMan probe into the magnetic compound, carrying out fluorescent quantitative PCR amplification, and monitoring a fluorescent
signal in real time to obtain a Ct value; establishing a
standard curve between the
oxytetracycline concentration and the Ct value of the fluorescent quantitative PCR, and determining the oxytetracycline concentration in the sample to be detected according to the
standard curve. Based on an indirect competitive mechanism, through
signal conversion design, combination of oxytetracycline and the
aptamer is converted into quantitative analysis of free sDNA, and through integration of three technologies of aptamer free recognition,
magnetic bead targeted capture and qPCR
signal amplification, the sensitivity and anti-interference capability of oxytetracycline detection are improved.