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Doxycycline detection kit and preparation method thereof

A detection kit and doxycycline technology, which is applied in the field of determination of biological immunological methods, can solve the problems of large error in results, long analysis time, and time-consuming, etc., and achieve the effects of rapid response, simple operation, and convenient use

Inactive Publication Date: 2011-10-12
北京库尔科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography and liquid chromatography-mass spectrometry have high sensitivity and reliable results, but the determination methods are cumbersome, time-consuming, and costly, and operators need professional training, which is difficult to popularize; microbial methods are simple to operate, but have poor specificity and detection The cycle is longer and the result error is larger
Enzyme-linked immunosorbent assay is a competitive immunoassay method using enzyme markers, which has the advantages of high sensitivity, large detection capacity, and low cost. 1-4 hours), still has certain limitations

Method used

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  • Doxycycline detection kit and preparation method thereof
  • Doxycycline detection kit and preparation method thereof
  • Doxycycline detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: the preparation of doxycycline detection kit

[0033] 1. Preparation of doxycycline-conjugated antigen

[0034] Coupling doxycycline with bovine serum albumin to synthesize a doxycycline-bovine serum albumin conjugate as a doxycycline-conjugated antigen.

[0035] 2. Preparation of anti-doxycycline monoclonal antibody

[0036] BALB / C mice were immunized with doxycycline-conjugated antigens as immunogens, and hybridoma cell lines secreting anti-doxycycline monoclonal antibodies were obtained through hybridoma technology; antibodies were produced in large quantities by the method of inducing ascites in vivo, and Protein G was used The column was purified to obtain anti-doxycycline monoclonal antibody.

[0037] 3. Preparation of colloidal gold

[0038] Take 100ml of 0.01% chloroauric acid aqueous solution and heat to boil. Quickly add 1ml of 1% trisodium citrate aqueous solution as needed, continue to boil for about 5min, orange-red appears. The colloidal...

Embodiment 2

[0047] Example 2: Application Evaluation of Doxycycline Detection Kit

[0048] 1. Conformity rate of negative reference product

[0049] Use phosphate buffered solution (PBS: 0.01mol / L, pH 7.5) to prepare a standard solution of chloramphenicol with a concentration of 100ng / mL for 10 parallel tests. Observe the test results after 10 minutes. Both T and C lines show red bands. The result was negative.

[0050] Use phosphate buffer solution (PBS: 0.01mol / L, pH 7.5) to prepare a standard solution of streptomycin with a concentration of 100ng / mL for 10 parallel tests. Observe the test results after 10 minutes. Both T and C lines show red bands. The result was negative.

[0051] 2. Conformity rate of positive reference products

[0052] Use phosphate buffer solution (PBS: 0.01mol / L, pH 7.5) to prepare doxycycline standard substances with concentrations of 15, 30, and 45ng / mL, and carry out 10 parallel tests for each concentration, observe the test results after 10 minutes, and th...

Embodiment 3

[0059] Embodiment 3: the detection method of doxycycline detection kit

[0060] 1. Sample pretreatment

[0061] Serum: extract the serum, centrifuge or let it stand, and take the transparent supernatant for use; if the serum is excessively hemolyzed, dilute the serum with distilled water for one time before performing the experiment, otherwise the test piece will be too dark red, which will affect the test results.

[0062] Urine: Test directly with urine. If the urine is visible and turbid, it needs to be centrifuged, filtered, or the supernatant should be taken after precipitation for testing.

[0063] Milk: Centrifuge at 5000r / min at 4°C for 15min, remove the upper layer of fat and then detect.

[0064] Tissue sample: take 5-10g of tissue sample and mash it, add 20-30ml (0.01mol / L, pH 7.5) PBS, incubate at 80°C for 30min, bathe in water for 10min, centrifuge at 4°C, 5000r / min for 15min, remove the upper layer of fat , take the supernatant for detection.

[0065] Feed: Ta...

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Abstract

The invention belongs to the technical field of detection of biological immunological methods, and particularly relates to a doxycycline detection kit and a preparation method thereof. The kit comprises a sample pad (1), a colloidal gold pad (2), a nitrocellulose membrane (3), a sample suction pad (4) and a poly vinyl chloride (PVC) support plate (5); the colloidal gold pad is anti-doxycycline colloidal gold labeled monoclonal antibody glass fiber (or non-woven fabric); and a doxycycline coupling antigen serving as a detection line (T line) and a goat-anti-mouse antibody IgG serving as a quality control line (C line) are sequentially coated on the nitrocellulose membrane. The doxycycline detection kit is prepared by adopting a colloidal gold immunochromatography assay technology; and the preparation method is simple, can be used for detecting the doxycycline probably existing in a sample, and has the characteristics of convenience in use, simplicity in operation, rapid reaction, economy, practicality and the like.

Description

technical field [0001] The invention relates to the technical field of determination of biological immunological methods, in particular to a detection kit for rapid detection of doxycycline by colloidal gold immunochromatography and a preparation method thereof. Background technique [0002] Doxycycline, also known as deoxyoxytetracycline, also known as doxycycline, is a tetracycline drug, and its properties are light yellow or yellow crystalline powder, smelly, and bitter. Soluble in water or methanol, slightly soluble in ethanol or acetone, insoluble in chloroform. It is mainly used for upper respiratory tract infection, flat strip inflammation, biliary tract infection, lymphadenitis, cellulitis, senile chronic bronchitis, etc. caused by sensitive gram-positive bacteria and gram-negative bacilli. It is also used to treat typhus, Qiang insect disease, mycoplasma pneumonia, etc. It can still be used to treat cholera, and also to prevent falciparum malaria and leptospirosis...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/544G01N33/558
Inventor 李峰陈立柱
Owner 北京库尔科技有限公司
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