1. General

T-2 toxin is a mycotoxin produced by various Fusarium. It mainly pollutes wheat, barley, corn and

other food crops and their products, which constitute a great hazard to human health and animal

husbandry. T2 mainly affects the function of the blood, liver, kidney, pancreatic muscle and

lymphocyte. The clinical symptoms of T2 toxin poisoning is anorexia, vomiting, diarrhea, growth

retardation, reproductive and neurological dysfunction etc.The use of Trichothecenes(T2) ELISA

test kit to analyze T2 residue of samples is fast and accurate.

This kit is a new generation of fungal toxins detection products in application of ELISA technology

and the operation time is shorter than 60min, to minimize operating errors and work intensity.

2. Principle

This test kit is based on the competitive enzyme immunoassay for the detection of T2 in the sample. The coupling antigens are pre-coated on the micro-well stripes. The T2 in the sample and the

coupling antigens pre-coated on the micro-well stripes compete for the anti-T2 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 T2 in it. This value is compared to the

standard curve and the T2 concentration is subsequently obtained.

3. Application

This test kit can be used to detect T2 in corn, rice, wheat, beans, peanuts, oats, feed and other

samples qualitatively and quantitatively.

4. Technical specifications

Sensitivity: 0.2 ppb

Detection limit:

Peanuts, corn, oats, beans, feed etc. …………………………………………………24ppb

Wheat……………………………………………………………………………………..20ppb

Recovery rate:

110±15%

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), 2 ppb, 6 ppb, 18 ppb, 54 ppb, 162 ppb

Standard working concentration: 0ppb, 0.2ppb, 0.6ppb, 1.8ppb, 5.4ppb, 16.2ppb

3) T2 Enzyme conjugate ……………………………………………………………… 6ml

4) T2 Antibody working solution……………………………………………………… 6 ml

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

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

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

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

6. Materials required but not provided

1) Equipment: microtiter plate spectrophotometer (450 nm/630nm), oscillator, vortex shaker,

centrifuge, balance: 0.01g quantity sensitive, gaduated pipettes: 25ml, ear wash ball, funnel,

triangle bottle with plug, polystyrene centrifuge tube: 10ml,50ml, quantitative analysis of filter paper

2) Micropipettes: single-channel 10~100µl, 200~1000µl, multi-channel 250µl

3) Reagents: methanol（analysis pure）, deionized water

7. Sample pre-treatment

Solution preparation before sample pre-treatment:

1) washing buffer

Use 1 part of concentrated washing buffer（10×） and dissolve with 9 parts of deionized water to

obtain the ready to use washing buffer. Solution is stable for 1 month when stored at 4 °C.

2) sample extract solution

Use 3 part of methanol and dissolve with 2 parts of deionized water to obtain the ready to use

sample extract.

3) 6 T2 standards

Take six 1.5ml centrifuge tubes, numbered from 1 to 6. Take six concentrations of standard (10 ×)

(0ppb, 2ppb, 6ppb, 18ppb, 54ppb, 162ppb) 50μl ,add to the above six tubes, then each tube adding 450μl deionized water to dilute them by 1: 9, obtain 0ppb, 0.2ppb, 0.6ppb, 1.8ppb, 5.4ppb, 16.2ppb the standard solution(see the following table). Configured standard solution should be used in one week (4 ℃ storage)

Samples Preparation

7.1 Preparation of Peanuts, corn, oats, beans, feed etc.

− weigh 1g of ground sample into a suitable container and add 20 ml sample extract solution

− shake vigorously for five minutes (manually or with shaker)

− filter the extract through filter

− dilute 1 part of the obtained filtrate with 5 part of distilled water or deionized water.

− take the diluted liquid to test

Dilution factor: 120

7.2 Preparation of wheat sample

− weigh 1g of ground sample into a suitable container and add 20ml of sample extract solution

− shake vigorously for 5 minutes (manually or with shaker)

− filter the extract through filter

− take 1ml supernatant, add 2ml methylene dichloride, shake, centrifuge for 5min at 4000r/min,

take methylene dichloride layer, blow to dryness at 50℃.

− add 5 ml 10% methanol, shake evenly.

− take the liquid to test

Dilution factor: 100