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A. Jehanli- ISEI-10th Symposium-2011

A. Jehanli, J. Dunbar & S. Skelhorn IPRO Interactive Ltd. Wallingford, Oxfordshire, U.K. Development and validation of an oral fluid collection device and its use in the immunoassay of salivary steroids and immunoglobulins in sports persons. A. Jehanli- ISEI-10th Symposium-2011.

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A. Jehanli- ISEI-10th Symposium-2011

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  1. A. Jehanli, J. Dunbar & S. Skelhorn IPRO Interactive Ltd. Wallingford, Oxfordshire, U.K. Development and validation of an oral fluid collection device and its use in the immunoassay of salivary steroids and immunoglobulins in sports persons A. Jehanli- ISEI-10th Symposium-2011

  2. IPRO represents “individual profiling” Established at Wallingford, Oxfordshire in September 2009 Main business: the development of rapid real-time / near-side quantitative tests for biological markers relevant to the sports world. Examples: Salivary IgA, IgG, Cortisol, Testosterone and DHEA The Company A. Jehanli- ISEI-10th Symposium-2011

  3. Ease of collection, no need for specialist staff Unlimited supply. A normal person produces more than 1 litre per day Non-invasive / intrusive Drugs in saliva not bound to proteins. Advantages of saliva testing A. Jehanli- ISEI-10th Symposium-2011

  4. “not nice” socially unacceptable Compliance issues: Time, food & drink Mouth full of bacteria - immediate freezing required Freeze-thaw cycles & centrifugation required to break down mucins and allow manipulation Methods of saliva collection Expectoration: drooling or spitting Advantage: provides neat sample, no dilution Disadvantages: A. Jehanli- ISEI-10th Symposium-2011

  5. Release of analytes from swab material Need for centrifugation Sample preservation Methods of saliva collection Swab material: cotton, polyester, nylon polymer (Rayon) etc Advantages: socially acceptable, quick, volume controlled by swab size Disadvantages: A. Jehanli- ISEI-10th Symposium-2011

  6. Buffer acts to extract or release analyte from swab Buffer can preserve the sample No need for centrifugation Methods of saliva collection Swab material & Buffer: Advantages: Disadvantage: No sample volume determination A. Jehanli- ISEI-10th Symposium-2011

  7. Commercially available collection devices M. Gorschl (2008) Clin Chem. 54, 1759 A. Jehanli- ISEI-10th Symposium-2011

  8. Develop a simple and quick to use saliva collection device Allows rapid collection time Provides sufficient known volume for both laboratory and real-time analysis Provides good recovery of analytes by using a “universal” extraction buffer Aim A. Jehanli- ISEI-10th Symposium-2011

  9. The IPRO OFC (oral fluid collection) device A. Jehanli- ISEI-10th Symposium-2011

  10. The IPRO OFC procedure Before After Volume Indicator A. Jehanli- ISEI-10th Symposium-2011

  11. Collection analysis Subjects: 57 male, 28 female Age range:17-82 A. Jehanli- ISEI-10th Symposium-2011

  12. Collection Time A. Jehanli- ISEI-10th Symposium-2011

  13. Collection Volume A. Jehanli- ISEI-10th Symposium-2011

  14. Analyte recovery from device vs neat saliva determined using in-house & Salimetrics ELISA test kits IgA & IgG recovery over 90% Cortisol and DHEA recovery over 85% Testosterone recovery more variable but over 70% Analyte Recovery A. Jehanli- ISEI-10th Symposium-2011

  15. IPRO OFC is a simple and rapid means of collecting saliva with adequate analyte recovery, for immediate or subsequent analysis Compliance in challenging sporting environments greatly enhanced when using the OFC as opposed to passive drooling Conclusion / Practical Application A. Jehanli- ISEI-10th Symposium-2011

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