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Kit and method for detecting beta-lactam antibiotic and tetracycline antibiotic

A technology of tetracyclines and tetracycline drugs, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity, high detection cost, and high requirements for detection personnel, and achieve the effect of improving detection sensitivity and high sensitivity

Active Publication Date: 2013-05-15
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microbial obstruction detection methods mainly include the inhibition zone test, turbidity test, and color-changing detection gold standard kit method. This method has low detection cost and low price, but it is low in sensitivity, takes a long time, and the existence of drug-resistant bacteria easily leads to false detection; physical and chemical The detection method is currently the main method for detecting β-lactam and tetracycline antibiotics, including liquid chromatography, chromatography / mass spectrometry, gas chromatography, etc., with high accuracy, but its detection cost is high and the equipment is complicated , Higher requirements for testing personnel, long testing time and other disadvantages. At present, the commonly used immunological analysis methods are mainly enzyme-linked immunoassay and radioimmunoassay. The required testing costs are still high, which is not suitable for low-cost enterprises. detection needs
[0004] Chinese invention patent 200810162588.5 (CN101429241A) discloses "the method and application of penicillin and carrier protein coupling product and β-lactam penicillin antibody preparation", the ELISA kit prepared by the penicillin monoclonal antibody in this method can only To detect three β-lactam antibiotics, penicillin G, ampicillin and carbenicillin, the detection sensitivity of the prepared colloidal gold test strip is low, and there are few types of detection drugs
At present, there is a lack of technology and application in the field of immunoassays that can simultaneously detect β-lactams and tetracyclines.

Method used

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  • Kit and method for detecting beta-lactam antibiotic and tetracycline antibiotic
  • Kit and method for detecting beta-lactam antibiotic and tetracycline antibiotic
  • Kit and method for detecting beta-lactam antibiotic and tetracycline antibiotic

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, the constitution of the kit that detects β-lactam class and tetracycline class antibiotic

[0059] 1. Freeze-dried microwell strips with gold-labeled antibodies

[0060] Microwell reagents are housed in the microwell strip, wherein 1 is a microwell strip, and 2 is the micropore of freeze-dried cephalosporin drug-specific antibody-colloidal gold marker and tetracycline drug-specific antibody-colloidal gold marker Reagent;

[0061] The micropore strip that contains micropore reagent has micropore plug 3 ( figure 2 with image 3 );

[0062] The micropore reagent of the freeze-dried cephalosporin drug monoclonal antibody-colloidal gold marker and the tetracycline drug monoclonal antibody-colloidal gold marker in the microwell strip, the cephalosporin drug specific antibody-colloidal gold marker The lyophilized amount is 0.20-0.25 μg / ml; the lyophilized amount of tetracycline drug-specific antibody-colloidal gold marker is 0.30-0.40 μg / ml.

[0063] Two, t...

Embodiment 2

[0070] The preparation method of kit described in embodiment 2, embodiment 1

[0071] 1. Preparation of kits

[0072] The preparation method of the kit mainly includes the following steps:

[0073] 1) Prepare freeze-dried microwell reagents of cephalosporin monoclonal antibody-colloidal gold marker and tetracycline monoclonal antibody-colloidal gold marker in a microwell strip, and cover the microwell strip with a microwell plug.

[0074] 2) Prepare a detection line (B) coated with a cephalosporin drug-carrier protein conjugate, a detection line (T) coated with a tetracycline drug hapten-carrier protein conjugate, and a detection line (T) coated with a goat anti-mouse anti-antibody The reaction membrane of the quality control line (C);

[0075] 3) Assemble the reaction film prepared in 2) with the sample absorbent pad, water absorbent pad, protective film, and bottom plate into a test strip;

[0076] 4) 1) and 3) prepared freeze-dried microporous reagents, test strips, micr...

Embodiment 3

[0135] Embodiment 3, the application of test strip

[0136] One, detect β-lactam antibiotics and tetracycline antibiotics in milk with the kit described in Example 1

[0137] The kit of the invention can detect milk samples. The general detection method is as follows:

[0138] 1. Detection method

[0139] Remove the microwell plug covered on the microwell strip, and drop the sample solution to be detected into the microwell strip containing the microwell reagent to obtain a mixed solution 11 of the microwell reagent and sample solution, mix well, and incubate 5min, insert the end of the test strip 12 with the MAX line mark down into the microwell strip, and watch the result within 5min, as Figure 7 shown.

[0140] 2. Judgment of test results

[0141] When the concentration of β-lactam antibiotics or (and) tetracyclines in the sample is higher than or equal to the minimum detection limit of the kit, the colloidal gold antibody will combine with β-lactam antibiotics or (an...

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Abstract

The invention discloses a kit and a method for detecting beta-lactam antibiotic and tetracycline antibiotic. The kit comprises micropore strips, micropore reagents, test paper and micropore plugs, wherein the micropore reagents are lyophilized colloidal gold labeled cephalosporins and tetracyclines specific antibodies, the cephalosporins specific antibodies comprise cephalosporins monoclonal antibodies and cephalosporins polyclonal antibodies, the tetracyclines specific antibodies comprise tetracyclines monoclonal antibodies and cephalosporins polyclonal antibodies; the test paper comprises a sample absorption pad, a reaction film, a water absorption pad, a protection film and a base plate which are connected in sequence, the reaction film comprises two detection lines and a quality control line, each detection lines comprises cephalosporins-carrier protein conjugates and tetracyclines hapten-carrier protein conjugates, and the quality control line comprises antiantibodies. According to the invention, the method for detecting beta-lactam antibiotic and tetracycline antibiotic by using the kit is simple, rapid, visual, accurate, portable, wide in application range, low in cost and easy to popularize.

Description

technical field [0001] The invention relates to a kit and method for detecting β-lactam and tetracycline antibiotics. Background technique [0002] Antibiotics are one of the most important medical discoveries in the 20th century and have played a huge role in controlling human infectious diseases. However, in recent years, the increase of drug-resistant pathogens caused by the abuse of antibiotics not only increases medical expenses, but also increases the morbidity and mortality of infectious diseases. In addition, the widespread use of antibiotics in agriculture and animal husbandry is also a non-negligible cause of drug resistance. Antibiotics such as β-lactams and tetracyclines are commonly used in the dairy industry to treat livestock mastitis, metritis and other diseases. Some producers also add antibiotics to feed to improve the feed conversion rate of livestock and poultry. According to the investigation of the US Food and Drug Administration (FDA), inattention to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577
Inventor 何方洋万宇平吴鹏罗晓琴崔海峰冯静冯月君余厚美陈炜玲
Owner BEIJING KWINBON BIOTECH
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