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Test strip for testing ochratoxin A and application of test strip

An ochratoxin and test strip technology, which is applied in the field of test strips for detecting ochratoxin A, can solve the problems of on-site screening of unpleasant samples, long sample preparation time, expensive instruments and equipment, etc., and achieves short detection time and low cost. , the effect of the simple method

Active Publication Date: 2015-04-29
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are liquid chromatography, gas chromatography, mass spectrometry and capillary electrophoresis methods for detecting OA in grain and feed. These methods have high sensitivity and stable results, but the required equipment is expensive, sample preparation takes a long time, and is not suitable for a large number of samples. On-site screening limits its application

Method used

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  • Test strip for testing ochratoxin A and application of test strip
  • Test strip for testing ochratoxin A and application of test strip
  • Test strip for testing ochratoxin A and application of test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 Preparation of ochratoxin A detection test strip

[0041] The preparation method of this test strip mainly comprises the following steps:

[0042] 1) Prepare a conjugate release pad sprayed with ochratoxin A monoclonal antibody-colloidal gold marker;

[0043] 2) Prepare a reaction membrane with a detection line coated with ochratoxin A hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;

[0044] 3) Assemble the conjugate release pad prepared in 1) and 2), the reaction membrane, the sample absorption pad, the water absorption pad and the PVC bottom plate into a test strip.

[0045] The following step-by-step detailed description:

[0046] 1. Preparation of ochratoxin A hapten

[0047] Add 40mg of ochratoxin, 30mg of p-phenylenediamine and a small amount of DMAP into 5ml of dry DMF. Slowly add the mixture of 40mg of DCC in 1ml of dry DMF dropwise at 0°C. After the dropwise addition, naturally warm up to ro...

Embodiment 2

[0080] The detection of ochratoxin A residue in the sample of embodiment 2

[0081] 1. Sample pretreatment

[0082] Weigh 3.0±0.05g crushed feed sample into a 15ml or 50ml polystyrene centrifuge tube; add 6ml of acetonitrile, cover the bottle tightly, shake for 5min; room temperature (20-25℃), centrifuge at 3000g or more for 5min; pipette 1ml Put the supernatant into a glass centrifuge tube and dry it in a water bath at 50-60°C under nitrogen flow; add 0.4ml of the sample complex solution, vortex for 30s, and wait for detection.

[0083] 2. Test with test strips

[0084] Use a straw to absorb the sample solution to be tested and add 3 drops vertically to the sample hole, start timing when the liquid flows, react for 5-10 minutes, and judge the result.

[0085] 3. Analyze test results

[0086] Negative (-): Both the T line and the C line are colored, indicating that the concentration of ochratoxin A in the sample is lower than the detection limit, such as figure 2 (a).

[...

Embodiment 3

[0089] Example 3 sample detection example

[0090] 1. Detection limit test

[0091] Take blank grain and feed samples, add ochratoxin A to them respectively to a final concentration of 50, 100, and 200 ng / ml, take a test strip for detection, and repeat the measurement three times for each sample.

[0092] When using test strips to detect grain and feed samples, when the concentration of ochratoxin A in it is 50ng / ml, two red lines visible to the naked eye are displayed on the test strip, which is negative; when the concentration of ochratoxin A in it is When it is 100 or 200ng / ml, the quality control line of the test strip develops color, but the detection line does not develop color, which is positive, indicating that the detection limit of this test strip for ochratoxin A in grain and feed is 100ng / ml.

[0093] 2. False positive rate and false negative rate test

[0094] Take 20 positive samples of grain and feed with known ochratoxin A content greater than 100ng / ml and 20...

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Abstract

The invention discloses a test strip for testing ochratoxin A and application of the test strip. The test strip comprises a sample absorbing pad (1), a conjugate releasing pad (2), a reaction film (3), a water absorbing pad (4) and a bottom board (7), wherein a test line (5) coated with an ochratoxin A hapten-carrier protein conjugate, and a quality control line (6) coated with an anti-goat anti-mouse antibody are arranged on the reaction film, and the conjugate releasing pad (2) is sprayed an ochratoxin A monoclonal antibody- colloidal gold marker. The invention further provides a method for testing the ochratoxin A residues in grain and feed by applying the test strip for the ochratoxin A. The test strip provided by the invention has the characteristics of simple operation, high sensitivity, high testing speed, low cost and the like, and therefore, the test strip is suitable for the screening and the on-site supervision of a large number of samples.

Description

technical field [0001] The invention relates to a test strip for detecting ochratoxin A and its application, in particular to a colloidal gold test strip for detecting ochratoxin A, which is especially suitable for grains such as soybeans and corn and feed (raw materials, Detection of ochratoxin A residues in batches and concentrates). Background technique [0002] Ochratoxin A is produced by various Aspergillus and Penicillium species that grow on grains (wheat, corn, barley, oats, rye, rice, millet, etc.), peanuts, vegetables (beans), etc. The toxin can also be present in meat such as pigs and hens after animals have ingested moldy feed. Ochratoxin A mainly damages the liver and kidney of animals. This toxin mainly causes kidney damage, and a large amount of toxin may also cause intestinal mucosal inflammation and necrosis in animals. OA-producing bacteria are widely distributed in nature and pose a potential threat to human health and animal husbandry development. In ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/558G01N2333/38G01N2333/385
Inventor 万宇平何方洋冯才伟罗晓琴冯静赵正苗崔海峰余厚美
Owner BEIJING KWINBON BIOTECH
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