- 原理
側流體免疫層析法 (lateral flow immunochromatographic assay)，為快篩試劑的使用原理，藉由抗體抗原間的專一性與免疫親和力做檢測，被普遍應用在、病毒感染確認、妊娠測試、警局臨檢驗毒等方面。此方法多用於定性檢測，且顯色標定物上也有許多不同的種類，目前大多數的顯色方式是利用抗體吸附在膠體金（colloid gold）來呈色。
新月毒素(群)（trichothecenes）依照化學結構式的不同，可分成A、B、C、D四型:
1️⃣A型新月毒素➡️T-2 毒素(T-2 toxin, T-2) 、HT-2 毒素(HT-2 toxin, HT-2)
2️⃣B型新月毒素➡️嘔吐毒素(deoxynivalenol, DON)與雪腐鐮刀菌烯醇(nivalenol, NIV)
3️⃣C型新月毒素➡️巴豆素(crotocin)、燕茜毒素(baccharin)
4️⃣D型新月毒素➡️黑葡萄穗菌毒素(satratoxin)、漆斑菌素(roridin)
⚠️其中A、B、C型新月毒素皆由真菌(Fusarium spp.)產生⚠️
⚠️食品中污染物質及毒素衛生標準，尚未包含T-2 毒素、HT-2 毒素⚠️
↪️T-2毒素、HT-2毒素常見於穀物產品(如小麥、大麥、玉米、燕麥、米、黑麥及大豆...等)，當T-2毒素進入動物體內之後，也可能被代謝成HT-2毒素等多種代謝產物。
↪️此二種毒素含有乙醯基（acetyl group）的結構，此種結構會降低動物體內粒線體電子傳遞鏈的活性，而形成過多的自由基 (free radicals)，繼而造成脂質過氧化，並影響動物細胞膜之完整性，最後造成細胞壞死(necrosis)
- 結果判讀
極限:
偵測極限(limit of detection, LOD)： 50 ppb
定量極限(limit of quantification, LOQ)：50 ppb
偵測範圍:50–500 ppb(樣品超過500ppb需重新稀釋及測量)
- 樣品準備(SAMPLE PREPARATION)
The sample to be tested should be collected according to accepted sampling techniques (see FGIS sampling protocol or contact your Neogen representative). Obtain a representative sample (minimum 100 g). Grind the sample so at least 95% of the ground material passes through a 20 mesh sieve (about the particle size of fine espresso).
- 樣品萃取(SAMPLE EXTRACTION)
1. Weigh out 10 g ± 0.1 g of sample into extraction cup.
2. Add contents of one (1) MAX 1 aqueous extraction packet to the extraction cup.
3. Add 50 mL distilled or deionized water to the extraction cup.
4. Vigorously shake, using hand or mechanical means, for 3 minutes, or blend for 1 minute.
5. Allow the sample to settle, then filter with a filter syringe or Whatman #4 filter pa per to collect a minimum of 3 mL filtrate into a sample collection tube. Or, you may also pipette 1 mL of sample into a 2.0 mL micro-centrifuge tube, and centrifuge for 30 seconds using a microcentrifuge (approx. 2,000 x g).
- 操作流程(TEST PROCEDURE)— Raptor Integrated Analysis Platform
1. Place the appropriate number of red sample dilution cups into a sample cup rack. Label cups if necessary.
2. Add 100 µL of sample extract to each red sample dilution cup.
3. Add 1,500 µL of green-labeled sample diluent to red sample dilution cup. Mix by pipetting up and down five times.
4. Fully insert a Reveal Q+ MAX for T-2/HT-2 test strip into a Raptor cartridge.
5. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader.
6. Enter Sample ID if desired.
7. Add 400 µL of sample extract from the red sample dilution cup to the Raptor cartridge.
a. The Raptor system will start automatically.
b. Additional samples can be started in the other ports while the first sample is processing.
8. Results will be displayed on the Raptor screen after the 5-minute testing period is complete.
- 稀釋步驟(Dilution Procedure) — Raptor Integrated Analysis Platform(樣品超過500ppb需重新稀釋及測量)
1. Add 100 µL sample filtrate to a sample collection tube.
2. Add 400 µL distilled or deionized water to the sample collection tube. Mix well by pipetting up and down five times, or cap the tube and shake.
3. Add 100 µL of diluted sample extract (from step 2) to each red sample dilution cup.
4. Add 1,500 µL of sample diluent (green labeled bottle) to the red dilution cup with sample extract. Mix by pipetting up and down 5 times.
5. Insert the Raptor cartridge containing the test strip into any of the three ports within the Raptor.
a. The bar code on the test strip will be read. The system identifies the type of test strip and the lot number. If the lot number is not found in the system, the bar code reader in the front of the Raptor will turn on automatically.
b. Scan the QR code found on the tube containing the test strips. The information will be stored on the reader.
6. Enter Sample ID if desired.
7. Add 400 µL of sample extract from the red sample dilution cup to the Raptor cartridge.
a. The Raptor system will start automatically.
b. Additional samples can be started in the other ports while the first sample is processing.
8. Results will be displayed on the Raptor screen after the 5-minute testing period is complete.
NOTE: The Raptor reader will not calculate your dilution. Final result displayed will need to be multiplied by 5.
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