1. 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

2. 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

3. 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

4. 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

5. 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

6. 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

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

8. 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

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

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

11. 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

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

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

14. 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

15. 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