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Fluorescence detection test strip for simultaneously detecting four types of pathogenic bacteria, and preparation method and application thereof

A technology for fluorescence detection and pathogenic bacteria, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of high cost, low detection efficiency, low sensitivity, etc., and achieve the effect of low cost, low detection cost, and improved detection sensitivity.

Inactive Publication Date: 2019-04-09
YANGTZE NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For above-mentioned deficiencies in the prior art, the object of the present invention is to provide a kind of fluorescence detection test strip that detects 4 kinds of pathogenic bacteria simultaneously and its preparation method and application, solve the existing sensitivity of the detection method of foodborne pathogenic bacteria Problems of low cost, high cost and low detection efficiency

Method used

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  • Fluorescence detection test strip for simultaneously detecting four types of pathogenic bacteria, and preparation method and application thereof
  • Fluorescence detection test strip for simultaneously detecting four types of pathogenic bacteria, and preparation method and application thereof
  • Fluorescence detection test strip for simultaneously detecting four types of pathogenic bacteria, and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 A kind of fluorescence detection test strip that detects 4 kinds of pathogenic bacteria simultaneously

[0041] This embodiment provides a fluorescence detection test strip for four kinds of pathogenic bacteria based on upconversion nanomarker-aptamer recognition, including a PVC base plate 1, a sample pad 2, and an aptamer-functionalized upconversion nanoprobe binding pad 3 , a nitrocellulose membrane 4 and an absorbent pad 7; four detection lines 5 and quality control lines 6 are sequentially arranged on the nitrocellulose membrane 4; the adjacent pastings are connected and stacked.

[0042] The binding pad (3) is coated with an upconversion nanoparticle-pathogen first nucleic acid aptamer complex; the upconversion nanoparticle is an avidin-modified upconversion nanomaterial; the first nucleic acid The aptamer is capable of specifically recognizing Escherichia coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes, and its...

Embodiment 2

[0060] Embodiment 2: A kind of preparation method of the fluorescence detection test strip that detects 4 kinds of pathogenic bacteria simultaneously

[0061] The preparation method of this test strip mainly comprises the following steps:

[0062] (1) Avidin-coupled amino-functionalized upconversion nanomaterials

[0063] Quantitatively weigh 10mg NaYF 4 , YB / Tm up-converting nanomaterials were dispersed in 5 mL of PBS (0.01 mol / L), ultrasonically dispersed for 15 min, and 1.25 mL of 25% glutaraldehyde solution was added. Then, the mixed solution was shaken and incubated at room temperature for 2 hours, centrifuged, and the supernatant was discarded to obtain a solid powder. Wash three times with 0.01mol / L PBS to remove excess glutaraldehyde, resuspend the powder in 4mL0.01mol / L PBS, ultrasonically disperse, add 1mg / mL avidin, shake and incubate at room temperature for 12h, centrifuge, discard Clear, wash the material several times, and dry the obtained precipitate at 37°C ...

Embodiment 3

[0074] Embodiment 3: the usage method of the fluorescence detection test strip that detects 4 kinds of pathogenic bacteria simultaneously

[0075] (1) Test strip detection

[0076] Take out the prepared test strip, insert the end of the glass fiber membrane into the sample solution to be tested, take it out after waiting for 10 minutes, and detect the fluorescent signals of T line and C line at 980nm excitation and 452nm emission wavelength.

[0077] (2) Result judgment

[0078] Positive: If any one or more of the quality control line 6 and the detection line 5 develop color, it means that the target bacteria are contained in the sample to be tested, and the sample is positive. For the result judgment, see image 3 shown.

[0079] Negative: If the test line 5 does not develop color, and only the quality control line 6 develops color, it means that the sample to be tested does not contain the four food-borne pathogenic bacteria, and the sample to be tested is negative. See ...

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Abstract

The invention discloses a fluorescence detection test strip for simultaneously detecting four types of pathogenic bacteria, and a preparation method and application thereof. The bottom layer of the test strip is a supporting plate; the supporting plate is pasted with a sample cushion, a combination cushion, a nitrocellulose membrane and an adsorption cushion in sequence, wherein the sample cushion, the combination cushion, the nitrocellulose membrane and the adsorption cushion are tightly connected; the nitrocellulose membrane is provided with four detection lines and one quality control linein sequence; the combination cushion is coated with an upconversion nanoparticle-pathogenic bacteria first aptamer compound; the detection line is independently coated with a pathogenic bacteria second aptamer-streptavidin compound, i.e., a capture probe; and the quality control line is coated with a polyT-sequence-streptavidin compound, i.e., a quality control probe. The test strip has the advantages of high sensitivity, good specificity and low cost, can realize the simultaneous detection of four types of pathogenic bacteria in a complex sample, and has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a fluorescent detection test strip for simultaneously detecting four kinds of pathogenic bacteria, a preparation method and application thereof. Background technique [0002] Foodborne diseases induced by foodborne pathogens are one of the most prominent food safety problems in the world today. According to the information released by the World Health Organization, there are hundreds of millions of cases of foodborne diseases in the world every year, even in developed countries, at least 1 / 3 of the people suffer from foodborne diseases. [0003] At present, the identification and detection methods for foodborne pathogens are mainly based on three principles: 1) microbial culture method; 2) specific DNA hybridization; 3) antigen-antibody recognition technology. Among them, the traditional microbial culture method is time-consuming and inefficient, and cannot detect ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/558G01N33/56911G01N33/56916G01N33/56938
Inventor 卢春霞李昌满高晓旭
Owner YANGTZE NORMAL UNIVERSITY
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