Method and detection test strip for on-site rapid detection of bovine-derived and duck-derived components

A detection reagent and specific technology, which can be used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., and can solve the problem of lack of rapid detection products.

Active Publication Date: 2020-05-12
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no product for on-site rapid detection of bovine-derived and duck-derived ingredients

Method used

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  • Method and detection test strip for on-site rapid detection of bovine-derived and duck-derived components
  • Method and detection test strip for on-site rapid detection of bovine-derived and duck-derived components
  • Method and detection test strip for on-site rapid detection of bovine-derived and duck-derived components

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The following examples are used to further illustrate the present invention, but should not be interpreted as limitation of the present invention, under the premise of not departing from the spirit and essence of the present invention, the modification or embellishment of the present invention all belong to the scope of the present invention. Specific detection of source components in the sample of Example 1

[0039] Taking beef products and duck products as objects, the RPA amplification reaction was carried out after DNA extraction, and the test strips were used for detection.

[0040] 1. Sample preparation and storage

[0041] 1.1 Sampling

[0042] Collect 1 g each of beef products and duck products, and store them at -20°C for later use.

[0043] 1.2 DNA template preparation

[0044] The DNA template was prepared using the commonly used phenol-chloroform crude extraction method (Sambrook J, Fritsch E F, Mani Artis T. Molecular Cloning Experiment Guide [M]. 2nd Ed...

Embodiment 2

[0063] Embodiment 2 sensitivity test

[0064] Using 0.01ng, 0.1ng, 1ng, 10ng, and 100ng of bovine and duck DNA as templates, amplification was carried out according to the conditions of Example 1. The result is as image 3 and Figure 4 shown. image 3 Bovine DNA is used as a template, and SEQ ID NO 1 to SEQ ID NO 3 is used as primers and probes for detection. When the template quality is 0.1ng, a purple band can still appear at the T line of the test strip. Figure 4 Duck DNA is used as a template, and SEQ ID NO 4 to SEQ ID NO 6 are used as primers and probes for detection. When the template quality is 0.1ng, a purple band can still appear at the T line of the test strip, indicating high sensitivity.

Embodiment 3

[0065] Embodiment 3 cattle and duck mixed sample detection

[0066] Duck DNA with a mass fraction of 50%, 25%, and 5% was mixed into bovine DNA as a mixed template, and amplified according to the conditions of Implementation 1, and the amplified products were detected by test strips. The result is as Figure 5 shown. Bovine and duck-derived components can be detected in all mixed samples.

[0067] Embodiment 4 test strip

[0068] The invention provides a colloidal gold test strip for detecting RPA nucleic acid amplification products, which includes a gold label pad, a detection line and a quality control line, and the gold label pad is adsorbed with an anti-FITC gold label antibody. The coating concentration of the gold-labeled antibody is (1 mg / L) and the coating volume is (1 μ L); the detection line is adsorbed with biotin antibody, and its coating concentration is (1 mg / L) and the coating volume (1 μ L ); the quality control line is adsorbed with the secondary antibody...

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Abstract

The invention provides a method and a kit for on-site rapid detection of bovine-derived and duck-derived components, and belongs to the technical field of animal provenance component molecular detection. The kit provided by the invention comprises a combination of bovine and duck specific recombinase polymerase amplification (RPA) labeled primers and probes, and the sequences of the combination are shown as SEQ ID No.1-SEQ ID No.3 and SEQ ID No.4-SEQ ID No.6. The kit provided by the invention also comprises a detection test strip which can be used for carrying out multiple detection on RPA amplification products. When the combination of RPA primers and probes is adopted to amplify DNA of a sample, whether the sample contains bovine and duck-derived components or not can be judged by detecting amplification products by the test strip. The detection method provided by the invention has the advantages of high sensitivity, strong specificity, no need of special equipment and reaction timeof only 30 minutes. The invention provides an effective, accurate and reliable means for detection of bovine-derived and duck-derived components, has the characteristics of simplicity, convenience, rapidness and strong specificity, and is suitable for on-site detection.

Description

technical field [0001] The invention relates to the field of animal molecular biology, in particular to a test strip for on-site rapid detection of bovine-derived and duck-derived components. Background technique [0002] With the gradual improvement of people's living standards, people's consumption of meat is increasing day by day, and the price difference of meat is also gradually widening. It is more and more common for low-priced meat to pass off as high-priced meat. At present, in the production and sales of beef products in the domestic and foreign markets, incidents of cheating consumers by means of adulteration and shoddy appear frequently. Among them, counterfeiting beef with cheap duck meat is currently the main means of adulteration. This kind of adulteration not only involves issues such as nutritional value and food safety, but also seriously violates the legitimate rights and interests of consumers. Therefore, the safety of the source of meat products has be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6844C12Q1/6804C12N15/11
CPCC12Q1/6804C12Q1/6844C12Q1/6888C12Q2521/507C12Q2522/101C12Q2565/625
Inventor 刘榜付明周翔张庆德张全旺
Owner HUAZHONG AGRI UNIV
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