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Kit and method for detecting beta-lactamase

A lactamase and kit technology, applied in chemical instruments and methods, measuring devices, instruments, etc., can solve problems such as being unsuitable for on-site use, high requirements for testing personnel, prone to false positives, etc. Sensitivity of response, wide range of effects

Active Publication Date: 2012-05-16
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used detection methods for β-lactamase mainly include liquid phase method and microbial method. Although the liquid phase method has high accuracy, it has disadvantages such as complicated operation, high requirements for testing personnel, long detection time, and high cost. Microbial detection The cost of the method is low, but the detection sensitivity is low, false positives are prone to occur, and it takes a long time, so it is not suitable for on-site use

Method used

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  • Kit and method for detecting beta-lactamase
  • Kit and method for detecting beta-lactamase
  • Kit and method for detecting beta-lactamase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the composition of the test kit that detects β-lactamase

[0040] 1. Microporous Reagents

[0041] The micropore reagent has a micropore plug;

[0042] The colloidal gold-labeled cephalosporin monoclonal antibody is lyophilized on the micropore reagent, and the coating amount is 0.15-0.20 μg / ml.

[0043] 2. Enzyme test reagent

[0044] The micropore plate of the enzyme testing reagent has a micropore plug;

[0045] The penicillin drug is freeze-dried on the microwell plate of the enzyme test reagent, and the coating amount is 0.15-0.20 μg / ml.

[0046] Two, test paper (called test strip) ( figure 1 ):

[0047] The test paper is composed of a bottom plate, a sample absorption pad, a reaction film, a water absorption pad, and a protective film;

[0048] The sample absorbent pad 1, reaction film 2, water absorbent pad 3 and protective film are pasted on the bottom plate 6 in sequence, the end of the sample absorbent pad is connected to the reaction film...

Embodiment 2

[0054] The preparation method of kit described in embodiment 2, embodiment 1

[0055] 1. Preparation of kits

[0056] The preparation method of the kit mainly includes the following steps:

[0057] 1) Prepare freeze-dried microporous reagent plate with cephalosporin monoclonal antibody-colloidal gold marker;

[0058] 2) prepare freeze-dried enzyme test reagent with penicillin medicine;

[0059] 3) preparing a reaction membrane with a detection area coated with a cephalosporin-carrier protein conjugate and a quality control area coated with a goat anti-mouse anti-antibody;

[0060] 4) Assemble the reaction film prepared in 3) with the sample absorbent pad, water absorbent pad, protective film, and bottom plate into a test paper;

[0061] 5) Assemble the microwell reagent, enzyme test reagent and test paper prepared in 1), 2) and 3) into a kit.

[0062] The following step-by-step detailed description:

[0063] (1) Preparation of each component

[0064] 1. Synthesis and ide...

Embodiment 3

[0106] Embodiment 3, the application of test paper

[0107] One, detect β-lactamase in milk with the kit described in embodiment 1

[0108] The kit of the invention can detect milk samples. The general detection method is as follows:

[0109] 1) Detection method

[0110] Add dropwise the sample solution to be tested into the wells of the enzyme-labeled reagent plate, mix well, incubate at 45°C for 25 minutes, warm up at room temperature for 10 minutes, add 200 μl of the enzyme test reagent after warming to the microwell reagent plate dropwise, and mix well , Insert the end of the test paper marked with the MAX line down into the well of the microwell reagent plate, and watch the result within 5 minutes.

[0111] 2) Result judgment

[0112] When the concentration of β-lactamase in the sample is higher than or equal to the minimum detection limit of the kit, β-lactamase decomposes the penicillin drug in the enzyme test reagent, and the colloidal gold antibody cannot combine ...

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Abstract

The invention discloses a kit and a method for detecting beta-lactamase. The kit comprises a microporous reagent, an enzyme test reagent and test paper, wherein a cephems medicament specificity-colloidal gold marker is freeze-dried in the microporous reagent; a penicillin medicament is freeze-dried in the enzyme test reagent; a cephems medicament specificity antibody is a cephems medicament monoclonal antibody or a cephems medicament polyclonal antibody; the test paper is formed by connecting a base plate, a sample absorption pad, a reaction film, a water absorption pad and a protection film sequentially; the reaction film comprises a detection region and a quality control region; the detection region is coated with cephems medicament and carrier protein conjugate; and the quality controlregion is coated with an antibody. The method for detecting the beta-lactamase by using the kit is simple, quick, intuitive and accurate, is wide in application range and low in cost, and is easy to popularize and use.

Description

technical field [0001] The invention relates to a kit and method for detecting β-lactamase. Background technique [0002] With the improvement of people's living standards, the drug residues in food of animal origin have been paid more and more attention by the international community. β-lactam antibiotics are the earliest discovered and most widely used antibiotics, and the most reported residues in milk and animal tissues. β-lactam antibiotics refer to a class of antibiotics containing a β-lactam ring in their chemical structure, mainly including penicillins and cephalosporins, and their function is characterized by being able to inhibit the activity of bacterial mucopeptide transpeptidase, thereby preventing bacterial Synthesis of cell walls, exhibits bactericidal activity. In animal production, β-lactam antibiotics are widely used to control mastitis in dairy cows, and treat animal urinary tract, gastrointestinal tract and respiratory tract infections. However, due to...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/532C07K16/44
Inventor 何方洋万宇平冯才伟冯静汪善良罗晓琴何丽霞赵正苗冯才茂崔海峰李勇王建霞
Owner BEIJING KWINBON BIOTECH
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