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Special test box for enzyme-linked immunosorbent assay of class-B trichothecenes

A trichothecenes, enzyme-linked immunosorbent technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of high detection cost, complicated processing, unfavorable promotion and application, etc., and achieve rapid detection, low price, Easy-to-use effects

Inactive Publication Date: 2014-05-14
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the detection methods of class B trichothecenes include high performance liquid chromatography (HPLC), gas chromatography (GC), thin layer chromatography (TLC), etc., because the sample pretreatment of the above analytical methods is relatively complicated , special technical personnel are required for operation, and the detection cost is expensive, which is not conducive to popularization and application

Method used

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  • Special test box for enzyme-linked immunosorbent assay of class-B trichothecenes
  • Special test box for enzyme-linked immunosorbent assay of class-B trichothecenes
  • Special test box for enzyme-linked immunosorbent assay of class-B trichothecenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The coated plate is coated with trichothecene type B solid-phase antigen, which adopts 96 or 48 or 24 well microwell plate, and uses 10 mmol / L pH9 Na2CO3—NaHCO 3 Dilute Class B trichothecenes-ovalbumin (OVA) to 1.0 μg / mL, add 100 μL to 96 or 48 or 24-well microplates, and place overnight at 8°C Or incubate in a 37°C oven for 2 h, discard the coating solution, wash 3 times, add 5% bovine serum albumin (dissolved in pH 7.5 phosphate buffer), block overnight at 2°C or place in a 37°C oven for 1.5 h, discard the blocking solution, blow dry, seal the strip and store it in a freezer at -2°C.

[0034] The class B trichothecene standard substance is represented by a standard substance of deoxynivalenol (DON), which is obtained by diluting the pure product of deoxynivalenol (DON), and the diluent is deionized water, DON Concentration: 0.001 ng / mL.

[0035] Preparation of Class B trichothecenes-bovine serum albumin (BSA) conjugates: Dissolve Class B trichothecenes in pyridine, ...

Embodiment 2

[0040] The coated plate is coated with the B trichothecene solid-phase antigen, which adopts 96 or 48 or 24-well microwell plate, and uses 100 mmol / L pH10 Na 2 CO 3 —NaHCO 3 Dilute Class B trichothecenes-ovalbumin (OVA) to 0.1 μg / mL, add 100 μL each to 96 or 48 or 24-well microplates, and place overnight at 5°C Or incubate in a 37°C oven for 2 h, discard the coating solution, wash twice, add 1% bovine serum albumin (dissolved in pH 7 phosphate buffer), block overnight at 8°C or place in a 37°C oven for 1.5 h , discard the blocking solution, blow dry, seal the strips and store in a -4°C refrigerator.

[0041] The class B trichothecene standard substance is represented by a standard substance of deoxynivalenol (DON), which is obtained by diluting the pure product of deoxynivalenol (DON), and the diluent is deionized water, DON Concentration: 1000 ng / mL.

[0042] Preparation of Class B trichothecenes-bovine serum albumin (BSA) conjugates: Dissolve Class B trichothecenes in py...

Embodiment 3

[0047] The coated plate is coated with trichothecene type B solid-phase antigen, which adopts 96 or 48 or 24-well microplate, and uses 55 mmol / L pH9.5 Na2CO 3 —NaHCO 3 Dilute Class B trichothecenes-ovalbumin (OVA) to 4.95 μg / mL, add 100 μL each to 96 or 48 or 24-well microplates, and place overnight at 2°C Or incubate in a 37°C oven for 2 hours, discard the coating solution, wash 5 times, add 3% bovine serum albumin (dissolved in pH 8 phosphate buffer), block overnight at 5°C or place in a 37°C oven for 1.5 h, Discard the blocking solution, blow dry, seal the strips and store in a refrigerator at 4°C.

[0048] The class B trichothecene standard substance is represented by a standard substance of deoxynivalenol (DON), which is obtained by diluting the pure product of deoxynivalenol (DON), and the diluent is deionized water, DON Concentration: 500 ng / mL.

[0049] Preparation of Class B trichothecenes-bovine serum albumin (BSA) conjugates: Dissolve Class B trichothecenes in py...

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Abstract

The invention discloses a special test box for the enzyme-linked immunosorbent assay of class-B trichothecenes, which has the advantages of fast, sensitive and accurate detection, quantification, simplicity in operation, low requirement on the sample purity and strong specificity and is particularly suitable for mass samples. Therefore, the invention also provides a preparation and detection method of the special test box. The special test box is characterized by comprising a concentrated washing solution, a concentrated complex solution, a color developing solution A, a color developing solution B, a stop solution, a coating board coating the solid-phase antigen against class-B trichothecenes, a standard product of class-B trichothecenes and an antibody freeze-dried product of class-B trichothecenes. After the detection, an absorbance value is detected at 450nm, and the content of class-B trichothecenes in the sample is calculated from a standard curve.

Description

[0001] technical field [0002] The invention relates to the technical field of biological and food safety detection, in particular to a special test box for ELISA detection of class B trichothecene toxins and a preparation and detection method. Background technique [0003] Class B trichothecenes are a class of metabolites produced by Fusarium fungi, which are widely distributed in nature, mostly in wheat, barley, corn and other grain seeds, and are the most dangerous food that occurs naturally Pollutants are very harmful to humans and animals. It not only causes acute and chronic poisoning of humans and animals, but also has potential risks of carcinogenicity, teratogenicity and mutagenesis. Type B trichothecenes mainly include deoxynivalenol (deoxynivalenol, DON), 3-acetyl deoxynivalenol (3-acetyl-DON, 3-Ac-DON), 15- Acetyl deoxynivalenol (15-acetyl-DON, 15-Ac-DON), nivalenol (NIV) and fusarenon X (FUS-X), etc. Its chemical structure is shown in the figure below (I) an...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531
CPCG01N33/543G01N33/531G01N2333/37
Inventor 杜道林吴湾湾
Owner JIANGSU WISE SCI & TECH DEV
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