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i-screen KHV Kit Economical Testers
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i-screen KHV Kit

A Brief Introduction to the i-screen™ KHV Kit:

The i-screen™ KHV Kit is one of the most sensitive viral detection tools for Koi, and is widely recognized as having the same sensitivity as Nested PCR. You can use it without being professionally trained, with the whole testing process taking around 3 hours, after which you can to obtain a highly accurate test result. The danger of KHV infection can therefore be minimized to a great extent, and the owner can always control possible virus infection beforehand.

Specifications:

  • Packing specification: 180 reactions.
  • Detection Limit: 10 copies per reaction.
  • Quantitative capability: 3 different levels of infection can be identified.
  • Sampling rate: for 40 specimens, only 3.5 to 4 hours from specimen preparation to the reveal of final results.
  • Built-in internal control system: eliminate false negative results from failed extraction or reaction.
  • Quantified positive standard: monitoring sensitivity of detection.

    • When to use:

    1. Quarantine before you place new fish into the main pond.
    2. After sudden change of temperature.
    3. After transporting the fish for a long time or distance.
    4. Suspected KHV-infected fish.
    5. Protection and quarantine before and after competition.

    I. Components of the doses:


    Plastic bag: It contains Q-tip, Dropper, Spotting Tip(x 2), Detection Stick, Filter-Tip(x 2) and Regular-Tip.

    Aluminum bag: It contains Tube 2K (x 2), Tube 2-1, and Tube 0, 1, 3, 4, 5, 5-1, 6, 7 each.
    If any of the following is found, please keep the kits and avoid using them. Contact sales person immediately and ask for exchange.

    1. When the grinder is broken
    2. When the detection stick is loosen or detached
    3. The tips are broken
    4. Liquid leaked from Tube Nos. 0, 1, 2-1, 3, 4, 5, 5-1, 6, or 7
    5. Aluminum bag is inflated
    (We will not offer exchange service should the seals are opened on either the plastic or the aluminum bag)

    Storage condition:
    Always store i-screen™ kits in a freezer. Please avoid thawing and refreezing repeatedly. Use them before expiry date.

    II. i-screen™ Starter Kit:



    Tube Racks:
    The racks are plastic, in 2 different colors. There are 2 green racks and 4 red ones. There are 24 holes for you to place 1.5ml to 2ml tubes, and 15 holes for 0.5ml tubes on each rack. They can be stored from -30°C to 90°C, and are acid, alkaline, and salt proof.
    ĦiApplication MethodĦjUse for placing tubes during tests only.
    ĦiStorage ConditionĦjRoom temperature
    ĦiMaintenanceĦjWash them with water and keep them dried after being used.

    Reaction Oven:
    (Temperature default is fixed in our factory. Please do not re-adjust the default by yourself.)
    The reaction oven is made of stainless steel inside and painted outside on the rust-proof steel plate. Oven temperature can be adjusted by pushing the button, from room temperature to 80°C.

    ĦiSpecial attention for installationĦj
    1. Before you plug it in, always make sure the voltage on the socket is correct, or the oven will be damaged seriously, and you will also be in danger.
    2. When using the oven, place it horizontally on a fixed position (such as a table) and avoid shaking it.
    3. When using, the oven generates heat. Do not put anything on it so as to avoid accidents.
    ĦiApplication methodĦj
    1. Plug it into a socket and switch it on. When its indicator light is on, it means the power supply is working correctly.
    2. Check temperature on the display to see whether pre-set 60°C (minus or plus 2°C is acceptable) is reached after 30 minutes. The oven can be used when it has reached the operating temperature of 60°C.
    3. Contact your sales person if the temperature inside the oven cannot reach 60°C (again, minus or plus 2°C is acceptable) after pre-heating for 30 minutes.
    4. It is strongly suggested that you do not adjust temperature on the display settings, as this may lead to test failures.
    ĦiMaintenanceĦj Unplug and cover the oven, if you want to leave it unused for a longer period, to protect it from dust and water.

    ĦiCleaning methodĦj
    1. Always unplug the oven before cleaning.
    2. Clean outer part and the inside of the oven with paper tower (preferably wet).
    3. Pay special attention when cleaning the inside of the oven. Never clean or touch the sensor at the left hand upper corner. Then, leave the door of the oven open until it is dried in room temperature.
    Micro Pipette:
    The light-weight, plastic, quantitative pipettes are designed for long-term use. Their accuracy will not be influenced by temperature and can be operated with only a single hand, to avoid contamination while changing their tips. There are 2 types of pipettes; one is black, and the other is white.

    ĦiApplication methodĦj
    1. The pipettes are a special instrument for testing. When you use it for the first time, you can practice sucking-in doses by using distilled water as a substitute, so as to familiarize yourself with its two-stop operating methods.
    2. Before liquid intake, make sure the tips are capped atop of the pipettes. (Regular-tip for Black Pipette and Filter-tip for White Pipette). Never under any circumstances should you reuse any tips. Please discard used tips immediately each test.
    ĦiMaintenanceĦj
    1. Make sure you clean the pipettes and store them in a proper place after each use.
    2. Never drop them or throw them around, or they will not function correctly.
    3. Do not heat them, and do not use them for liquid over 70°C.
    ĦiCleaning methodĦj
    1. Use wet paper tower to clean the pipettes.
    2. If you find that little amount of liquid is left in the top of the pipettes, contact your salesperson and ask for professional cleaning methods, or purchase new pipettes.

    III. Pipette Usage:

    Up-load tips:
    Use your finger tips to hold the widest part of tip. Slightly screw it onto the top of the pipette, and make sure tip is tightened.
    Remove tips:
    Press the reject bottom to remove the tip. Discard the tip immediately.
    		Pipette in:
    1. Press the plunger to its first stop.
    2. Immerse the tip in the liquid.
    3. Allow the plunger to move up smoothly.
    4. Move the pipette's tip out of the tube.
    		Pipette out:
    1. Place the pipette tip in the tube.
    2. Press the plunger to its first stop.
    3. Move the pipette tip out of the tube.
    4. Allow the plunger to move up smoothly.

    Attention:

    1. Each time before taking specimen: clean your both hands thoroughly with soap, and dry them with tissue paper, paying special attention to the fingertips.
    2. Pre-heat the Reaction Oven for at least 30 minutes.
    3. You are strongly recommended that you allocate three separate spaces for the whole testing process: dissolving area, sampling area, and reacting area.
    4. All components are for a single test, never reuse components. After detecting, all the tubes should be capped and discarded immediately.
    5. Please prepare bleach dilution in the reacting area. (Dilute bleach in a bottle with a sealed cap, e.g. Mineral water bottle.) This is for disposing by using the Spotting Tips and Tube 2.
    6. Please mark clearly on different specimens before you do the tests by using marker pen or stickers. (For Tube 0, 1, 2, and the Detection Stick.)
    7. Shake each tube so as to precipitate the reagent at the bottom of it every time before using.
    8. When using pipettes, press only to its First Stop. Please refer to "Pipette Usage" on the right.
    9. Please read carefully the "i-screen™ Instruction Manual" enclosed in the Starter Kit if you are going to use any equipment for the first time. Should you need any help, please do not hesitate to call the nearest distributor for further service and detailed explanation.
    The Dissolving Area

    Step 1
    Step 2
    1. Take out the test kits with doses equivalent to the number of specimens planned to test from the freezer. Unfreeze the kits.
    2. Swing Tube 2-1 gently to precipitate the liquid.
    3. Apply the Regular Tip to the Black Pipette and transfer the defrosted liquid from Tube 2-1 to Tube 2K at one time along the tube wall while transferring. Apply the same method to the second Tube 2K for dissolving.
    4. Leave the remaining liquid in Tube 2-1 and discard it.
    5. Remove the Regular Tip and discard it.
    1. Use the Dropper to transfer all of the defrosted liquid from Tube 5-1 along the tube wall to Tube 5. Cap Tube 5 and swing gently.
    2. Move all of the test kits to the Sampling Area.
    The Sampling Area

    *Please use the Q tip to scrape the mucus of the fish for 5 times in the red zone as indicated for collecting specimen.
    Step 3
    1. Follow the Sampling Instruction on the left for collecting the specimen required.
    2. Cut off the side with the mucus of the fish into Tube 0.
    3. Seal the tube with its cap, shake its until bubbles pop up inside. Wait for its submerge for 5 minutes.
    4. If there is more than one specimen, please mark the specimens properly on each cap of every test kit correspondingly so as to avoid mistaking.

    Step 4
    Step 5
    1. Swing Tube 1 to precipitate the liquid.
    2. Apply the Filter-Tip to the White Pipette and transfer upper liquid (without bubbles) from Tube 0 AT A TIME to Tube 1 by inserting its tip into the liquid surface.
    3. Remove the Filter-Tip and cap Tube 1 on. Flick it three times and swing it three times.
    4. To avoid contamination, please cap Tube 0 with any remaining specimen and discard it.
    1. Apply a new Filter-Tip on the White Pipette and transfer liquid from Tube 1 AT ONE TIME only by inserting its tip to Tube 2K. Do not release your thumb on the button until the tip is removed from the liquid surface. Cap Tube 2K on.
    2. Repeat the previous steps for another Tube 2K. Cap Tube 2K on.
    3. To avoid contamination, please cap Tube 1 with any remaining liquid and discard it.
    The Reacting Area

    Step 6
    Step 7
    1. Move all the remaining test kits to the Reacting Area.
    2. Place all Tube 2K on the red tube rack and place the rack in the Reaction oven which has been pre-heated for at least 30 minutes.
    3. Adjust the oven's timer to 90 minutes. Activate the timer and start its counting down.
    4. Note: In order to retain the temperature, avoid leaving the Reaction oven door unclosed for more than 10 seconds.
    5. Pour liquid bleach (around 3ml) into an empty bottle. Dilute with water 100 times so as to make its total volume 300ml. Screw the cap back on, and then it is ready for use.
    1. Take out the rack after 90 minutes.
    2. Prepare the Detection Stick and mark with specimen number on every stick c orrespondingly.
    3. Insert Spotting Tip under the liquid surface of Tube 2K and wait for 1 second. And then, touch the detection window of the Detection Stick for 1 second. Immediately discard the Spotting Tip and Tube 2K (with cap on) into the bottle of the bleach dilution.
    4. Repeat the previous steps for another Tube 2K with the second Spotting Tip. Do not overlap the two spotting areas.
    5. Screw the Detection Stick onto Tube 3 tightly. Repeat step 3 to 5 until all the tests are completed.

    Step 8
    Step 9
    1. Place all Tube 3 on the tube rack. Turn it over for 5 times, and then place them into the reaction oven for 5 minutes.
    2. After 5 minutes, transfer and screw the Detection Stick onto Tube 4. Submerge for 10 seconds under room temperature.
    1. Transfer and screw the Detection Stick onto Tube 5. Place all Tube 5 on the tube rack and turn it over for 5 times. Screw caps back on Tubes 3 and 4 and discard them immediately.
    2. Put the tube rack in the reaction oven for 5 minutes.
    3. Later, transfer and screw the Detection Stick onto Tube 6. Submerge for 10 seconds under room temperature.
    4. Cap Tube 5 and discard it.

    Step 10
    1. Transfer and screw the Detection Stick onto Tube 7. Place Tube 7 back on the tube rack and turn it over for 5 times. Put the tube rack in the reaction oven for 10 minutes.
    2. Retain Tube 6 with its cap on.
    3. After 10 minutes, return the Detection Stick to Tube 6 and submerge for 10 seconds under room temperature.
    4. Take out the Detection Stick and lay it horizontally on the tube rack. Put the detection window with its face up and dry it in the reacting oven for 10 minutes.
    5. Take out the Detection Stick for result interpretation.
    Result interpretation

    After detecting, it is strongly suggested that you discard the Detection Sticks within 24 hours. Do not bring the Detection Sticks out of the Reacting Area.

    Attention

    Due to the special nature of the KHV, positive result might not be shown during the latent phase. The actual meaning of "no KHV-infection" can be also interpreted as it is in the latent phase.

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