Rapid detection method for abrin toxin
A rapid detection method for abrin toxin is provided. The rapid detection method for abrin toxin includes the following steps: reacting an oligonucleotide chain substrate labeled with a fluorescent group and a quenching group, a buffer solution, a BSA solution, a to-be-detected sample and sterile water, incubating at a constant temperature, detecting a fluorescence signal value, and determining whether the abrin toxin exists based on a difference between a change in final fluorescence signal value and a change in final fluorescence signal value of a negative control; where the sample is judged as a positive sample when an average value of the detected final fluorescence signal values is greater than an average value of the final fluorescence signal values of the negative control by +3 times standard deviation. According to the detection method, the direct addition of the sample into the reaction system can reach the sensitivity of 30 ng/mL.
1. A rapid detection method for abrin toxin, comprising the following steps:
subjecting an adenine-containing oligonucleotide chain substrate labeled with a fluorescein (FAM) fluorescent group and a Black Hole Quencher 1 (BHQ1) quenching group, a buffer solution, a bovine serum albumin (BSA) solution, a to-be-detected sample, and sterile water to a reaction to obtain a product,
incubating the product at a constant temperature to obtain an incubated product,
detecting a fluorescence signal value 2-5 times to obtain final fluorescence signal values of the incubated product, and
determining whether the abrin toxin exists based on a difference between an average value of final fluorescence signal values of the incubated product and a final fluorescence signal value of a negative control; wherein
the buffer solution is an ammonium formate buffer solution, an ammonium acetate buffer solution, an ammonium citrate buffer solution, an ammonium acetate and ethylene diamine tetraacetic acid (EDTA) buffer solution, or an ammonium citrate and EDTA buffer solution;
the to-be-detected sample is enriched by using antibody-coated magnetic beads, wherein antibodies for coating the magnetic beads are specific for abrin toxin; and
the negative control is sterile water.
2. The rapid detection method according to claim 1 , wherein an amount of the adenine-containing oligonucleotide chain substrate labeled with the FAM fluorescent group and the BHQ1 quenching group, the buffer solution, the BSA solution, the to-be-detected sample, and the sterile water is 0.5-6 μL, respectively.
3. The rapid detection method according to claim 1 , wherein an amount of the antibodies for coating the magnetic beads is 20-30 μg per 0.5-2 mg of the magnetic beads.
4. The rapid detection method according to claim 1 , wherein the adenine-containing oligonucleotide chain substrate labeled with the FAM fluorescent group and the BHQ1 quenching group has a final concentration of 2-20 μM; the ammonium formate buffer solution, the ammonium acetate buffer solution, and the ammonium citrate buffer solution have a final concentration of 0.2-20 mM, respectively; in the ammonium acetate and EDTA buffer solution, ammonium acetate has a final concentration of 0.2-20 mM, and EDTA has a final concentration of 0.2-0.8 mM; in the ammonium citrate and EDTA buffer solution, ammonium citrate has a final concentration of 0.2-20 mM, and EDTA has a final concentration of 0.2-0.8 mM; and the BSA solution has a final concentration of 10-100 μg/mL.
5. The rapid detection method according to claim 1 , wherein the reaction is performed at a pH value of 3.5-4.8 and a temperature of 40-75° C. for 30-50 min.
6. The rapid detection method according to claim 1 , wherein the to-be-detected sample is judged as a positive sample when the average value of the final fluorescence signal values of the incubated product is greater than the final fluorescence signal value of the negative control by plus 3 times standard deviation.