▎ 摘　　要

NOVELTY - Preparing Datian acid immune detection composite probe involves adding biotin-labelled secondary antibody into the aqueous solution graphene incubating overnight at 4℃ to obtain complex I. The complex I obtained is centrifuged to remove the supernatant, then added with phosphate buffer saline (PBS) to re-disperse to obtain the complex II. The complex II is added with streptavidin labelled horseradish peroxidase into, incubated at 4℃ overnight to obtain complex III. The complex III is centrifuged to remove the supernatant, and then added with PBS buffer solution to re-disperse to obtain desired product. USE - Method for preparing Datian acid immune detection composite probe used in detecting Datian acid. ADVANTAGE - The prepared Datian acid immune detection composite probe helps in realizing the double-function multi-stage amplification of the colour signal, and improving the detection sensitivity. DETAILED DESCRIPTION - An INDEPENDENT CLAIM is included for a method of detecting Datian acid using immune detection composite probe, which involves: (a) adding 30 μL 50ng/mL large field soft sponge acid coating antigen in each micro-hole of the microarray chip, incubating for 2 hours at 37℃, removing the redundant large field soft sponge acid coating antigen, and then using PBST to wash the micropore for 3 times; (b) adding 40 μL bovine serum albumin (BSA) confining liquid with concentration of 5 wt.% in each micropore, incubating for 2 hours at 37℃ to close the active site, removing the redundant BSA confining liquid, and cleaning the micropore for 3 times with phosphate-buffered saline with Tween 20 (PBST); (c) adding large field acid standard product with different concentration of 15 μL in different micro-pores, then respectively adding 15 μL monoclonal antibody (1: 80000 dilution) in each micro-pore for competition reaction, mixing uniformly at 37℃ incubation for 2 hours, washing the micropore for 3 times with PBST; (d) adding 30 μL soft sponge acid immune detection composite probe solution (1: 600 dilution) in each micro-pore, 37℃ lower incubation for 2 hours, then using PBST to wash the micropore for 3 times; and (e) adding 30 μL 3,3',5,5'-Tetramethylbenzidine (TMB) chromogenic liquid in each micro-hole, using intelligent mobile phone to collect the chroma signal, according to the linear relation between each color intensity and sample concentration, calculating the concentration of the sample.