Lateral flow assay in Animal Disease Diagnosis


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Use of Rapid lateral flow assay in Veterinary Practice

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Lateral flow assay in Animal Disease Diagnosis

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