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Avian Influenza Virus Antigen Detection Tests. Mary Lea Killian USDA APHIS VS National Veterinary Services Laboratories Ames, Iowa. C. A. B. FluDetect ™ Test Strip Protocol. To tube provided: Add 8 drops extraction buffer. Add 200 μ l AAF (sample), mix gently

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avian influenza virus antigen detection tests

Avian Influenza VirusAntigen Detection Tests

Mary Lea Killian

USDA APHIS VS

National Veterinary Services Laboratories

Ames, Iowa

fludetect test strip protocol

C

A

B

FluDetect™ Test Strip Protocol

To tube provided:

  • Add 8 drops extraction buffer. Add 200 μl AAF (sample), mix gently
  • Place test strip directly into the sample so that the pink pad is just submerged into the extracted sample
  • Incubate strip for 15 minutes
binax now influenza a b test protocol
Binax NOW® Influenza A&B Test Protocol
  • Remove the device from the pouch just prior to testing—lay flat
  • Draw 100 μl sample into the pipet—make sure there are no air bubbles
  • Add the entire contents (drop by drop) to the white pad at the top of the strip
  • Peel off the adhesive liner, seal the device and incubate for 15 minutes
avian influenza antigen detection tests
Avian InfluenzaAntigen Detection Tests
  • Advantages
    • Rapid (15-20 minutes)
    • Highly specific (nucleoprotein target)
    • No special facilities required
  • Disadvantages
    • Expensive ($15-30/test)
    • Moderate sensitivity (70-80% compared to VI)
    • False positives (bacterial contamination)
    • Interference by blood (alkaline phosphatase)
avian influenza virus type a antigen test kit
Avian Influenza Virus Type A Antigen Test Kit
  • Recommended for testing sick/dead birds (high virus load)
  • FluDetect™–Conditional license for veterinary use
  • Test strips contain antibody to influenza type A nucleoprotein
  • FluDetect™: Tracheal, oropharyngeal, and cloacal swabs
  • Binax NOW®: Tracheal and oropharyngeal swabs
  • Confirmation of AI in hemagglutinating samples
viral transport media vtm
Preferred Media for use with FluDetect™

Brain-Heart Infusion (BHI) Broth, porcine origin

Tris-Buffered Tryptone Broth (TBTB)

Nutrient Broth (NB)

Peptone Broth (PB)

Viral Transport Media

Preferred Media for use with Binax NOW®

Amies Media

BHI broth

Dulbecco’s Medium

Hank’s Balanced Salt Solution (BSS)

Sucrose-Phosphate Buffer is NOT suitable for use with this test

Viral Transport Media (VTM)
interpretation of results fludetect
Interpretation of Results—FluDetect™

a POSITIVE sample is indicated by two pink/purple bands (control line and test line) that are clearly visible on the strip. The test line may be any intensity

Control band

Positive

Negative

A negative result indicates that no detectable influenza type A virus is present in the sample

interpretation of results binax now
Interpretation of Results—Binax NOW®

A NEGATIVE sample has a single pink/purple

control line in the bottom third of the window

A sample POSITVE for INFLUENZA A has a

pink/purple control line and a second pink/purple

line in the MIDDLE third of the window

A sample POSITIVE for INFLUENZA B has a

pink/purple control line and a second pink/purple

line in the TOP third of the window

The test is INVALID if the control

line remains BLUE or is not present

at all, whether a sample line exists

or not