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Method of detecting residue of small-molecule substance harmful to human body and a special kit

A technology of small molecular substances and immune reagents, which is applied in the field of detection of small molecular substance residues harmful to the human body, can solve the problems of long distance, and achieve the effects of long stable time, narrow emission spectrum and strong fluorescence

Inactive Publication Date: 2013-11-06
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, ELISA technology also has its own defects. Due to the specific combination of antigen and antibody, only one drug can be detected. Even if a broad-spectrum antibody is prepared, it can only realize the multi-residue detection of the same drug. The multi-drug residue testing of

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  • Method of detecting residue of small-molecule substance harmful to human body and a special kit
  • Method of detecting residue of small-molecule substance harmful to human body and a special kit
  • Method of detecting residue of small-molecule substance harmful to human body and a special kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, composition and preparation of kit

[0063] 1. An ELISA plate coated with avermectin, salinomycin and sulfaquinoxaline at the same time in the same well;

[0064] Abamectin coating was originally a conjugate of abamectin hapten and egg serum albumin.

[0065] Salinomycin coating was originally a conjugate of salinomycin hapten and ovalbumin.

[0066] The sulfaquinoxaline coating was originally a conjugate of sulfaquinoxaline hapten and human serum albumin (HSA).

[0067] 2. Luminescent compound:

[0068] 1) The conjugate formed by salinomycin monoclonal antibody and "quantum dot and goat anti-mouse antibody complex"; the characteristic emission wavelength of quantum dot is 585nm;

[0069] 2) The conjugate formed by "complex of abamectin monoclonal antibody and biotin" and "complex of avidin and quantum dot"; biotin is N-hydroxysuccinimide biotin (NHSB), The avidin is streptavidin; the characteristic emission wavelength of the quantum dot is 525nm;

[...

Embodiment 2

[0146] Embodiment 2, detection method

[0147] 1. Preparation of standard curve

[0148] Add 20 μl of salinomycin standard solution, 20 μl of avermectin standard solution, and 20 μl of sulfaquinoxaline standard solution to the same microwell of the ELISA plate coated with three kinds of coating materials, and then add three kinds of luminescence 40 μl of each complex was incubated at 37°C for 30 minutes, poured out the liquid in the well, washed 4 times with 260 μl / well, and patted dry. Set the excitation wavelength and emission wavelength (excitation wavelength 400nm, emission wavelength: the emission wavelength 585nm corresponding to the salinomycin luminescence complex; the emission wavelength 525nm corresponding to the abamectin complex; the emission wavelength corresponding to the sulfaquinoxaline luminescence complex 655nm;), read with a multi-functional microplate reader, and measure the relative fluorescence intensity.

[0149] Salinomycin standard solution is set at...

Embodiment 3

[0160] Embodiment 3, the effect of kit

[0161] (1) Sensitivity (IC 50 and detection limit)

[0162] The method for evaluating the reaction sensitivity of competitive enzyme-linked immunosorbent assay, commonly used is IC 50 Inhibitory concentration (referring to the drug concentration corresponding to 50% of the absorbance value of the standard solution) and detection limit, the lower the value of the two, the higher the sensitivity of the method. There are many definitions of the detection limit. The definition of the detection limit in this experiment is: the mean value of 20 blank samples or zero standard plus 3 times the standard deviation is the detection limit of the method.

[0163] According to the three standard curves in Experiment 2, the ICs corresponding to the three substances were obtained 50 Inhibitory concentration, as shown in Table 1-3.

[0164] 20 parts of milk samples not containing abamectin, salinomycin and sulfaquinoxaline were pretreated according ...

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Abstract

The invention discloses a method of detecting the residue of small-molecule substances harmful to the human body and a special kit. The special kit comprises a non-transparent micro-porous plate and a light-emitting compound, wherein each hole of the non-transparent micro-porous plate is filled with a coat antigen which is simultaneously coated with three kinds of small-molecule substances. The invention makes full use of the multi-color marking function of QDs, establishes a novel kit for simple and rapid detection of the residue and a method thereof, and realizes the multi-color marking through indirect marking of polyclonal antibodies and monoclonal antibodies in the veterinary drug by coupling the QDs with different particle sizes and targets with functional groups (such as an amino) with specific surfaces. The method comprises: obtaining quantum dots with different fluorescent characteristics through separation and purification, namely, multi-color antibody markers, using the multi-color antibody markers as fluorescent probes, and establishing a reaction system for synchronous analysis of various antigen components of different kinds, thereby realizing the synchronous detection of multiple kinds of residues of the veterinary drug in animal food. Moreover, the method has the advantages of simple operation, high fluorescence intensity and long stabilization time.

Description

technical field [0001] The invention relates to a method for detecting residues of small molecular substances harmful to human body and a special kit thereof. Background technique [0002] Vicious incidents of food quality and safety have occurred frequently in recent years, seriously threatening people's health and life safety. Food safety issues have become the focus of attention of the whole society. Effective monitoring of residues and hazardous substances such as veterinary drugs, pesticides, food additives, and hormones is the key to ensuring the safety of animal-sourced food and maintaining social stability. [0003] At present, the common method for detecting the above-mentioned residues or hazardous substances is enzyme-linked immunosorbent assay (ELISA). Its high sensitivity and specificity greatly simplifies the operation of residue analysis. Immunoassays are widely used in residue analysis and monitoring due to their low cost, rapidity, reliability, and high se...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/543G01N21/64
Inventor 丁双阳肖希龙沈建忠江海洋李建成夏曦王战辉陈军霞徐飞饶钦雄李金贵朱奎李存张启迪
Owner CHINA AGRI UNIV
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