1.  General

Deoxynivalenol, also known as Vomitoxin belongs to the trichothecene mold streptozotocin family, produced by some Fusarium fungi. Deoxynivalenol are presented in wheat, barley, corn and other grain seeds, the content is usually mg / kg (μg / mL, ppm) level. Because of their high cytotoxic and immunosuppressive properties, Deoxynivalenol constitutes a health threat to human and animal.

This kit is a new generation of fungal toxins detection products in application of ELISA technology, to minimize operating errors and work intensity.

2.  Principle

This test kit is based on the competitive enzyme immunoassay for the detection of Deoxynuvalenol in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Deoxynuvalenol in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti-Deoxynuvalenol antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Deoxynuvalenol in it. This value is compared to the standard curve and the Deoxynuvalenol concentration is subsequently obtained.

3.  Application

This test kit can be used to detect Deoxynuvalenol in corn and feed s corn, wheat, barley and animal feed samples qualitatively and quantitatively.

4.  Technical specifications

Sensitivity: 10 ppb

Detection limit:

Beer.............................................................................................. 10 ppb

Corn, peanut, wheat and other food crop, feed............................. .200 ppb

Recovery rate:

Beer ………………………………………………………………………..90±10%

Corn, peanut and other food crop, feed ………………………………..95±15%

Wheat, grist ………………………………………………………………..80±10%

Cross-reaction rate

Deoxynuvalenol ……………………………………………………………100%

Coefficient of variation:   <10%

5.  Components

1)  Micro-well strips: 12 strips with 8 removable wells each

2)  6 x concentrated standards ……………………………………………….1ml each

0 ppb (zero standard), 10 ppb, 20 ppb, 45 ppb, 135 ppb, 405 ppb

3)  Enzyme conjugate........................................................................................6ml

4)  Antibody working solution.............................................................................6ml

5)  Substrate A................................................................ ..................................6ml

6)  Substrate B............................................................................................ ......6ml

7)  Stop solution.................................................................................................6ml

8)  Concentrated washing buffer (10X)..............................................................40ml

9)  Concentrated sample diluent (10X)..............................................................40ml

6.  Materials required but not provided

1)  Equipment:  microtiter plate spectrophotometer (450 nm/630nm), vortex (oscillator optional), centrifuge (funnel optional), balance: 0.01g quantity sensitive, gaduated pipettes: 25 ml, polystyrene centrifuge tube: 50m(Triangle bottle with plug optional),Micropipettes: single-channel 20ml ~ 200ml, 200ml ~ 1000ml, multi-channel 250ml

2)  Reagents: deionized water

7.  Sample pre-treatment

Solution preparation before sample pre-treatment:

1)  washing buffer

Dilute the concentrated washing buffer at 1:9 with deionized water to use (1mL concentrated washing buffer + 9 mL deionized water). Solution is stable for 1 month when stored at 4 °C.

2)  Sample dilute (for food crops and feed)

Take out concentrated sample dilute from 2 ~ 8 ℃ environment, and return to room temperature. If there is a crystal separation, shock, or stirring to make it dissolved then use.

Dilute the concentrated sample diluent (10X) at 1:9 with deionized water to use (1mL concentrated sample dilute + 9 mL deionized water). Solution is stable for 1 month when stored at 4 °C.

Samples Preparation

Instructions (The following points must be dealt with before the pre-treatment)

1)       Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents;

2)       Before the experiment, each experimental utensil must be clean and should be re-cleaned if necessary, in order to avoid the contamination that interferes with the experimental results.

3)       Store un-handled samples in frozen.

4)       After-treatment samples store at 2-8 ℃ in the dark for 24h. All samples should be stored at

2-8 ℃ in the dark.

7.1 Preparation of beer

−Weight enough beer sample and remove CO2.

− Take 50 μl for assay directly.

Dilution factor: 1