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, J. Dunbar & S. Skelhorn IPRO Interactive Ltd. Wallingford, Oxfordshire, U.K. A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 The Company 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 A. Jehanli- ISEI-10th Symposium-2011
Advantages of saliva testing 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. A. Jehanli- ISEI-10th Symposium-2011
Methods of saliva collection Expectoration: drooling or spitting Advantage: provides neat sample, no dilution Disadvantages: “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 A. Jehanli- ISEI-10th Symposium-2011
Methods of saliva collection Swab material: cotton, polyester, nylon polymer (Rayon) etc Advantages: socially acceptable, quick, volume controlled by swab size Disadvantages: Release of analytes from swab material Need for centrifugation Sample preservation A. Jehanli- ISEI-10th Symposium-2011
Methods of saliva collection Swab material & Buffer: Advantages: Buffer acts to extract or release analyte from swab Buffer can preserve the sample No need for centrifugation Disadvantage: No sample volume determination A. Jehanli- ISEI-10th Symposium-2011
Commercially available collection devices M. Gorschl (2008) Clin Chem. 54, 1759 A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 Aim 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 A. Jehanli- ISEI-10th Symposium-2011
The IPRO OFC (oral fluid collection) device A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 The IPRO OFC procedure Before After Volume Indicator A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 Collection analysis Subjects: 57 male, 28 female Age range:17-82 Volume [mL] Time [seconds] Mean 0.53 33 SD 0.08 17.37 CV 15.03 53.38 Min 0.22 12.00 Max 0.64 102.00 A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 Collection Time A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 Collection Volume A. Jehanli- ISEI-10th Symposium-2011
A. Jehanli- ISEI-10th Symposium-2011 Analyte Recovery 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% A. Jehanli- ISEI-10th Symposium-2011
Conclusion / Practical Application 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 A. Jehanli- ISEI-10th Symposium-2011