Lateral flow assay in Animal Disease Diagnosis

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Use of Rapid lateral flow assay in Veterinary Practice

Use of Rapid lateral flow assay in Veterinary Practice

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  • 1. Bishor V. I Lateral Flow Immuno Assay in Disease Diagnosis Director, ubio Biotechnology Systems Pvt. Ltd, Kinfra Hi-tech Park, Kochi www.ubio.in
  • 2. Lateral flow assays
    • Rapid, Immunochromatography based.
    • Easy to do in the field.
      • Quick results in just a few minutes.
      • Require no instrumentation,
      • Allows decentralized implementation (should not require serum transport)
      • No, or low, capital expenditure.
      • Able to execute the test and read result in the field itself.
      • Room temperature storage, no refrigeration required.
  • 3. TYPICAL TEST STRUCTURE
  • 4.
    • The signal results from the formation of an immunocomplex on the membrane
    Y Detector Reagent Analyte Capture Reagent Membrane Chromogen Immunocomplex
  • 5. Assay Principle example
    • Eg Leptospira Antibody test in dogs
      • Test line: Pooled Leptospira antigen
      • Control line: Mouse IgG
      • Conjugate pad: Protein A colloidal gold
      • Sample pad: Buffered with Tris buffer.
  • 6. Test methodology
  • 7. Lateral flow assays in companion animal diagnosis
    • Canine distemper: antigen and antibody tests
        • Specimen: nasal or conjunctival swabs
    • Canine Parvovirus: Antigen test and antibody test
        • Specimen: fecal sample/serum
    • Canine Adeno virus: Antigen tests: Nasal swabs
    • Corona virus: antigen: Fecel swabs
    • Canine brucellosis: antibody tests
    • Ehrlichia canis: antibody test
    • Giardia : antigen in feacal sample
    • Heartworm : antigen: blood
    • Leptospira test: antigen and Ab tests
    • Feline tests: FeLV/FIV combo test
    Brucella/E.canis/ Ab tests CP/CD/Giardia test
  • 8. Vaccination titer check
    • Antibodies for CDV, CPV and Rabies can be assessed.
    • Color intensity of test line is
    • assessed for titer
  • 9. Lateral flow assays in Farm animal practice
    • Common tests are
    • Bovine Brucella Ab
    • Bovine TB Ab
    • FMD NSP Ab
    • Swine fever Ab test.
    • Rota virus test.
  • 10. Avian Tests
    • AIV Antigen test
    NDV Antigen test IBD Antigen test
  • 11. EXTARCTION
    • A rapid screening test which can detect Aflatoxin B1 in feed is very of critical importance in farms and feed manufacturing plants.
    • Lateral flow assays : using a competitive immunoassay format.
    • an easy and rapid extraction procedure using ethyl acetate.
    • Quantification from 2 to 100ppb.
    • This test is easier and cheaper compared to HPLC or TLC methods.
    • No need to handle toxic Aflatoxin standards and solvents.
    POSITIVE NEGATIVE Food testing and feed testing SEPARATION DILUTION TESTING
  • 12. Residue testing
    • Antibiotic residue testing in milk and meat.
    • Chloramphenecol in aquatic foods.
  • 13. Reagent Application Drying Process Lamination of Components Inspection/Marking Cutting Quality Control Assembly/Packaging SUMMARY OF MANUFACTURING PROCESSES
  • 14.
    • Reagent Application
    • Dispense: Test, control, conjugate
    • Coat: Conjugates, blocking, other pad treatments
    • Inspection/Marking: Dispensing defects
    • Drying Process
    • Objective is to reduce water content to less than 4 % level.
    • Used after reagent application processes.
    • Lamination of Components
    • Individual layers are bonded, using thin layers of adhesive.
    • Adhesive layers are usually prelaminated to plastic backing and cover layers.
    • Inspection/Marking
    • Lamination alignment defects
    SUMMARY OF MANUFACTURING PROCESSES
  • 15.
    • Cutting
    • Separate into individual test strips: 3-10 mm
    • Remove bad parts marked in inspection process
    • Quality Control
    • Sample testing of manufacturing lot for functionality
    • Assembly / Packaging
    • Seal test strip in foil pouch with desiccant, pipette, etc.
    SUMMARY OF MANUFACTURING PROCESSES
    • Manufacturing facility
    • Temperature and humidity controlled facility.
    • Relative Humidity below 25%
  • 16. Batch Manufacturing Equipment Line dispenser laminator Manual Dip Tray Guillotine Cutter Reel-to-reel web handling and dispensing platform Auto Laminator In-line manufacturing equipment
  • 17. Batch Process Line Layout T/C XYZ XYZ Conjugate Inspect/Mark Dip Tank Block Oven Dry Cutter Clamshell Clamshell Assemble Assembly Roller Assemble Assemble Seal Seal Seal Cut/Sort Inspect/Mark Laminate cover Laminate Master Strip Typ. 1.5 million tests limited by assembly & pouching 12-15 + operators Increase to 4-6 million by adding pouching and/or assembly machine Final QC
  • 18. New generation Lateral flow assay
    • Nucleic acid lateral flow assays: PCR amplified products can be detected.
    • Biochips: Multiplex microarray lateral flow format
    • Microfluidic chips: Lab on a chip: from sample extraction to detection without human intervention
    • Biosensors: electrochemical detection of immunoassays.
  • 19.
    • Thank you
    www.ubio.in