Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for detecting Fumonisin B1 based on magnetic enzyme-linked immunoassay

A technology for the detection of fumonisins and enzymes, which can be used in measurement devices, instruments, scientific instruments, etc., can solve problems such as unfavorable routine detection and use, and achieve the effects of high accuracy, strong specificity and high sensitivity

Active Publication Date: 2014-08-06
SHANGHAI JIAOTONG UNIV
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require strict pretreatment of test samples, expensive instruments such as high performance liquid chromatography, and professional operators, which are not conducive to on-site routine testing.

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
  • Method for detecting Fumonisin B1 based on magnetic enzyme-linked immunoassay
  • Method for detecting Fumonisin B1 based on magnetic enzyme-linked immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] Step 1, preparation of fumonisin B1 complete antigen (OVA-FB1)

[0033] Dissolve 2.5 mg of ovalbumin (OVA) in 0.1 ml of 0.01 M PB buffer, add 10 μl of 50% (V / V) glutaraldehyde (GA), and stir overnight at room temperature. Dialyze against PBS overnight at 4°C to remove excess GA. 0.5 mg of FB1 was dissolved in 0.2 ml of 25% (V / V) ethanol, added to activated OVA dialyzate (about 0.15 ml), added 0.1 ml of 1M carbonic acid buffer (pH 9.5), and stirred overnight at 4°C. Add 0.05ml 1M lysine (pH 7) and react at 4°C for 3h. Finally, dialyze with PBS for 72 hours, change the medium twice, and store at -20°C.

[0034] Step 2: Binding anti-FB1 monoclonal antibody to immunomagnetic beads

[0035] Take 50 μl of immunomagnetic beads, wash 3 times with washing solution (0.01M phosphate buffer, pH 7.4), then add binding buffer containing 5 μg of anti-FB1 monoclonal antibody (0.1M phosphate buffer, pH 8.2, containing 0.01% Tween-20) 200μl, shake at room temperature for 30 minutes. ...

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

No PUM Login to View More

Abstract

The invention relates to a method for detecting Fumonisin B1, which includes the following steps: coupling an FB1 hapten with an OVA to obtain a coupling object FB1-OVA; combining an FB1 monoclonal antibody with immunomagnetic beads; adding the antibody- magnetic bead combined object in an EP tube and washing; adding extract of a to-be-detected sample in the EP tube and oscillating; removing supernatant fluid and washing, adding OVA-FB1 in the EP tube and oscillating; removing supernatant fluid and washing, and adding the anti-FB1 monoclonal antibody into the EP tube, and oscillating; removing supernatant fluid and washing, and adding an HRP labeled goat-anti-mouse second antibody into the EP tube, and oscillating; removing supernatant fluid and washing, adding a luminous substrate and oscillating, and measuring the luminous value; and drawing a standard curve, and obtaining the corresponding concentration of FB1 through the standard curve and as per the luminous value of the to-be-detected sample. The method provided by the invention adopts the anti-FB1 monoclonal antibody, avoids cross reaction with mycotoxins in other grains, has strong specificity, and reduces the false positive rate.

Description

technical field [0001] The invention relates to a method in the technical field of biological detection engineering, in particular to a method for rapidly detecting fumonisin B1 in grains, grain-related products, and feed. Background technique [0002] Fumonisin B1 (Fumonisin B1, FB1) is a secondary metabolite produced by Fusarium moniliforme under certain humidity and temperature conditions, and it is widely distributed all over the world. In 1988, Gelderblom et al first isolated fumonisins from the culture solution of Fusarium moniliforme. Fumonisins can contaminate corn and its products, and have been detected in some grain-based products such as noodles, beer, condiments, and even in asparagus. Fumonisins have been reported to be associated with equine leukomalacia, pulmonary edema syndrome in pigs, liver cancer in rats and other diseases. In addition to the above-mentioned diseases, studies in the Transkei region of South Africa and the Linxian region of China have fo...

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
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
Inventor 严亚贤王元凯孙建和
Owner SHANGHAI JIAOTONG UNIV