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IDENTIFYING SNAKE SPECIES INVOLVED IN BITES

IDENTIFYING SNAKE SPECIES INVOLVED IN BITES. New Technology, New Options Disclaimer: I am not involved with any of the companies that make products covered in this presentation. ELISA (Enzyme-linked immunoassay). Inexpensive (relative! c. $275 for 3 reactions) relatively easy to perform

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IDENTIFYING SNAKE SPECIES INVOLVED IN BITES

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  1. IDENTIFYING SNAKE SPECIES INVOLVED IN BITES New Technology, New Options Disclaimer: I am not involved with any of the companies that make products covered in this presentation

  2. ELISA (Enzyme-linked immunoassay) • Inexpensive (relative! c. $275 for 3 reactions) • relatively easy to perform • fast (< 30 mins) • uses stable reagents • sensitivity of < 1 ng/ml

  3. ELISA (Enzyme-linked immunoassay) • False positives through low specificity/cross reactivity ESPECIALLY in regions with diverse snake faunas • False negatives through upper limit of detectability (c. 1 mg/ml in some spp) • first application to snakebite c. 35 years ago • Snake venom detection kit (SVDK) issued by CSL, Australia since 1988 is the still the only commercial diagnostic kit • Alternative biosensor approach advocated by Selvanayagam & Gopalakrishnakone in Toxicon in 1999.

  4. LABEL-FREE Kinetic profiling of immunotherapeutics Vaccine efficacy testing Eg. BLItzTM system Drop. Read. Done. Genius! http://www.blitzmenow.com

  5. 1. Pipette a 4 µL drop of sample into the sample holder. 2. Install a biosensor.   3. Select your assay and click GO. Results appear immediately!

  6. Disposable biosensor (can be re-used): c. $4.50/assay Non-denaturing, minimises non-specific binding Exceptional specificity even for crude media Bio-layer interferometry (BLI) Real-time data

  7. DNA diagnostics Is this what it takes?

  8. 1. Sample collection, storage and extraction Whatman™ FTA cards: Collect, archive, transport, and purify nucleic acids at room temperature minimal handling, streamlined, efficient workflow for sample collection and analysis. $3-5/sample (30 mins)

  9. 2. Biomarker amplification Ready-to-go PCR tubes: just add sample (c. $1.50/sample) Pre-dispensed, room temp stable, reliable and reproducible Custom formats including primers Also available in RT-PCR format for single-step amplification from RNA Still needs a conventional PCR machine (thermal cycler): c. $1700 PCR CYCLER: LavaAmp (prototype) 20-25 reactions max Powered by 4 x AA batteries or USB cable $300-$500, aiming for $100 Some development problems delaying it? lava-amp.com

  10. 3. Vizualisation: FlashGel® System (c. $430, plus c. $10/gel running costs) 5 minute separation and recovery TOTAL COST PER SAMPLE: $15-17 Equipment: c.£2000 (and can be used for other diagnostics as well http://dx.doi.org/10.1016/j.jip.2010.06.001

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