Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer

A technology of aflatoxin and test strips, applied in the field of detection, can solve the problems of unsuitability for on-site application and low sensitivity, and achieve the effects of simple and convenient detection time, high sensitivity, and avoiding interference

Active Publication Date: 2016-07-20
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
View PDF9 Cites 24 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a variety of detection methods have been established using the screened aptamers of AFB1 (which can also detect AFM1), such as colloidal gold colorimetry

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer
  • Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer
  • Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, detect the preparation of aflatoxin test paper strip

[0039] 1. Preparation of test strips for detecting aflatoxin

[0040] 1. Synthesis of detection probes and quality control probes

[0041] Fluorescein-labeled detection probes and biotin-labeled quality control probes were designed and synthesized, respectively.

[0042]The detection probe is aflatoxin B1 aptamer labeled with fluorescein, and the fluorescein is but not limited to Cy5.

[0043] The aflatoxin B1 aptamer can bind to the aflatoxin B1 hapten in the detection area, and the nucleotide sequence of the aflatoxin B1 aptamer is as follows:

[0044] 5'-Cy5-GTT-GGG-CAC-GTG-TTG-TCT-CTC-TGT-GTC-TCG-TGC-CCT-TCG-CTA-GGC-CCA-CA-3' (SEQ ID NO: 1)

[0045] The quality control sequence is a complementary single-stranded DNA molecule of a biotin-labeled aflatoxin B1 aptamer, which can be combined with the detection probe through base complementarity.

[0046] The nucleotide sequence of the quality contr...

Embodiment 2

[0067] Embodiment 2, test strip detects aflatoxin B1 or M1 sensitivity, linear range, specificity research

[0068] 1. Research on sensitivity and linear range of detection of aflatoxin B1

[0069] prepare image 3 The different concentrations of AFB1 (Fermentek Ltd, AF028) aqueous solutions shown were measured with the test strips prepared in Example 1, respectively.

[0070] The fluorescence intensity at the detection area and the quality control area was measured respectively with a test strip detection instrument ESEQuant-LR3. The result is as image 3 As shown, it can be seen that as the concentration of AFB1 increases, the fluorescence intensity at the detection area gradually decreases. Using the fluorescence intensity ratio of the detection area to the quality control area fluorescence intensity (Table 1), the standard curve for the detection of aflatoxin B1 is drawn as Figure 4 As shown, the results show that the sensitivity of the test strip is 0.5ng / mL; the lin...

Embodiment 3

[0081] Example 3, application of test strips in the detection of aflatoxin B1 in peanut meal

[0082] 1. Standard curve drawing

[0083] With the standard curve method in the detection of aflatoxin B1 sensitivity and linear range research in Example 1, the regression equation obtained is: y=-0.2076x+0.9881, R 2 =0.99, which is the unary regression equation of the aflatoxin B1 standard curve.

[0084] 2. Sample extraction

[0085] Select a sample of aflatoxin B1 in peanut meal (content 143.47ng / mL) determined by the national standard method (LC-MSMS), and use this test strip for determination. First weigh 2g of peanut meal into a 50mL centrifuge tube, add 8mL of n-hexane and 10mL of 70% methanol aqueous solution, shake for 5min, centrifuge at 4000 rpm at room temperature for 10min; remove the upper liquid, add 0.5mL of the lower liquid to 0.5mL of deionized water, Mix well, then take 0.5 mL of the mixed liquid and add 0.5 mL of 35% aqueous methanol solution, shake for 30 s; ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Linearaaaaaaaaaa
Linearaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a test strip for detecting aflatoxin B1 or M1 by utilizing an aptamer. The test strip for detecting the aflatoxin B1 or M1 by utilizing the aptamer, provided by the invention, comprises a sample absorption pad, a marker pad, a reaction film and a water absorption pad, wherein the marker pad is coated with a detection probe; the detection probe is an aflatoxin B1 aptamer marked by fluorescein; the nucleotide sequence of the aflatoxin B1 aptamer is sequence 1; the reaction film comprises a detection region and a quality control region; the detection region is coated with a conjugate formed by an aflatoxin B1 hapten and a carrier protein; and the quality control region is coated with a quality control probe, and the quality control probe is a conjugate formed by avidin conjugation probe marked aflatoxin B1 complementary single strand DNA (Deoxyribose Nucleic Acid)) molecules. The test strip for detecting the aflatoxin B1 or M1, provided by the invention, has the advantages of high sensitivity, accuracy in quantifying, strong specificity, simplicity and convenience, and short detection time.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a test strip for detecting aflatoxin B1 or M1 by using an aptamer. Background technique [0002] In recent years, excessive levels of aflatoxin in feed, milk, peanut oil, and grain have frequently appeared in some areas of my country, posing a serious threat to human health and causing great concern from all walks of life. Aflatoxin is a secondary metabolite produced by Aspergillus flavus and Aspergillus parasiticus, which has acute and chronic toxicity, mutagenicity, carcinogenicity and teratogenicity. Aflatoxins have a great impact on livestock and poultry, especially pigs are the most sensitive, mainly manifested as hepatomegaly, damage or cancer, intestinal bleeding, decreased immunity, poor growth and development, etc. Aspergillus toxin content shall not exceed 20ppb. Aflatoxin consists of about 20 similar chemical substances, mainly including aflatoxin B1, B2, M1, M2, G...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/533G01N33/558
CPCG01N33/533G01N33/558
Inventor 陈爱亮朱超张桂兰黄亚飞闫娇白文荟
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap