Vomitoxin嘔吐毒素(DON)-定量-快篩試紙-水溶性萃取-偵測儀器No. 9696

• 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。
  
  Vomitoxin嘔吐毒素，又稱脫氧雪腐鐮刀菌烯醇(deoxynivalenol, 簡稱DON)，屬於鐮刀黴菌毒素(新月毒素群)(trichothecenes)的一種，主要是由禾穀鐮孢菌(Fusarium graminearum)汙染大麥、小麥、玉米及相關的穀物飼料加工品所產生的天然毒素，DON最明顯的症狀是引起動物的嘔吐，此外，也會引起其他急性症狀，例如:腹痛、腹瀉、腸胃炎...等。
• 結果判讀
  極限:
  定量極限(limit of quantification, LOQ)：0.3 ppm
  偵測範圍:0.3–6 ppm(樣品超過6ppm需重新稀釋及測量)
   
• 樣品準備(Sample Preparation) — 10 g Sample
  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) — 10 g Sample
  1. Extract at a ratio of 1 part sample to 10 parts distilled or deionized water. For example, combine 10 g of ground sample with 100 mL of distilled or deionized water.
  2. Vigorously shake, using hand or mechanical means (250 rpm) for 3 minutes, or blend for 1 minute.
  3. Allow the sample to settle, then filter at least 4 mL with a filter syringe, or Whatman No. 1 filter paper.
  4. You may also pipette 1.5 mL of sample into a 2.0 mL microcentrifuge tube, and centrifuge for 30 seconds.
  5. The sample is now ready for testing.
   
• 操作流程(Test Procedure) — 10 g Sample
  1. Place the appropriate number of red sample dilution cups and clear sample 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 1,000 µL of sample diluent to red sample dilution cup. Mix by pipetting up and down 5 times.
  4. Transfer 100 µL of diluted sample extract into a new clear sample cup.
   
• 偵測儀器No. 9696操作(Reading a Test Strip)
  1. From the home screen: Select category: Mycotoxin Q+. Select test: Q+ DON Endpoint.
  2. Press run test. Select lot (if not listed, add the lot by scanning the QR code located on the device tube).
  3. Enter sample ID as needed — press accept.
  4. Fully insert the Reveal Q+ DON Endpoint test strip into the Neogen labeled cartridge adaptor with the sample end first.
  5. Insert cartridge into the track on the Solo reader — system will read automatically.
  6. Results are displayed on the unit. Samples greater than 6 ppm must be diluted and retested.
  7. To read the next test strip, press change at the bottom of the screen — this allows the user to enter the next sample ID. Press accept and insert cartridge containing the next test strip.
  Note: The Reveal Q+ for DON Endpoint can only be read using the Raptor Solo reader. Test strips should be read within 30 seconds of the completion of the 3-minute incubation.
   
• 稀釋步驟(Dilution Procedure)
  Samples greater than 6 ppm will need to be diluted and retested.
  1. Add 100 µL sample filtrate to a sample collection tube (use sample filtrate obtained in Sample Extraction Procedure from page 4, step 3 or 4).
  2. Add 500 µL distilled or deionized water to the sample collection tube. Mix well by pipetting up and down 5 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 1,000 µL of sample diluent (blue labeled bottle) to the red dilution cup with sample extract. Mix by pipetting up and down 5 times.
  5. Transfer 100 µL of diluted sample extract into a new clear sample cup.
  6. Place a new Reveal Q+ for DON test strip with the sample end down into the sample cup and set timer for 3 minutes. Ensure the test strip comes into contact with liquid and begins to wick.
  7. Remove the strip from the sample cup after it has developed for exactly 3 minutes. Immediately (within 30 seconds) read the test strip in the Raptor Solo reader.
  8. The final result displayed will need to be multiplied by 6.
  Note: The Raptor reader will not calculate your dilution. Final result displayed will need to be multiplied by 6. 
   
• FGIS方法-樣品萃取(FGIS Extraction Procedure) — 50 g Sample
  1. Weigh 50 ± 0.2 g of ground sample into a Whirl-Pak bag.
  2. Add 250 mL distilled or deionized water and close bag securely.
  3. Shake vigorously on a mechanical shaker at 250 rpm (or by hand with similar shaking action for 3 minutes.
  4. Allow the sample to settle
  5. Filter sample extract using a filter syringe to obtain 3-5 mL filtrate. (Note: filtrate should be clear of any particulates – if cloudy, re-filter.)
  6. Dilute the filtrate 3 +2 with distilled or deionized water.
  a. Example: add 900 µL filtrate(from step 5) to sample collection tube. Add 600 µL of distilled water. Mix well.
  7. Proceed to FGIS Test Procedure
   
• FGIS方法-操作流程(FGIS Test Procedure) — 50 g Sample
  1. Place the appropriate number of red sample dilution cups and clear sample cups into a sample cup rack. Label cups if necessary.
  2. Add 100 µL of diluted filtered extract (from step 6 of FGIS Extraction Procedure) to each red sample dilution cup.
  3. Add 1,000 µL of sample diluent to red sample dilution cup. Mix by pipetting up and down 5 times.
  4. Transfer 100 µL into a clear sample cup.
  5. Place a Reveal Q+ for DON test strip with the sample end down into the sample cup and set timer for 4 minutes. Ensure the test strip comes into contact with liquid and begins to wick.
  6. Remove the strip from the sample cup after it has developed for exactly 4 minutes. Immediately (within 30 seconds) read the test strip in the Raptor Solo reader using the Endpoint cartridge adaptor.
   
• FGIS方法-偵測儀器No. 9696操作(FGIS Reading a Test Strip) — 50 g Sample
  1. From the Raptor Solo home screen:
  a. Select category: Mycotoxin Q+.
  b. Select test: Q+DON Endpoint FGIS.
  2. Press run test. Select lot (if not listed, add the lot by scanning the QR code located on the enclosed QR code card).
  3. Enter sample ID as needed – press accept.
  4. Fully insert the Reveal Q+ DON Endpoint test strip into the Neogen labeled cartridge adaptor with the sample end first.
  5. Insert cartridge into the track on the Solo reader — system will read automatically.
  6. Results are displayed on the unit. Samples greater than 6 ppm must be diluted and retested.
  7. To read the next test strip, press change at the bottom of the screen — this allows the user to enter the next sample ID. Press accept and insert cartridge containing the next test strip.
  Note: The reveal Q+ for DON Endpoint can only be read using the Raptor Solo reader. Test strips should be read within 30 seconds of the completion of the 4-minute incubation.

• 產品說明書：連結
• 操作手冊：連結
• 物質安全資料表(Safety Data Sheet; SDS)：中文連結、英文連結

參考規範：食品中污染物質及毒素衛生標準