PTS Detect™ Cotinine System Training The PTS Detect™ cotinine system is the first and only point-of-care tobacco-use detection device offering quantitative results from a fingerstick. Cotinine, a metabolite of nicotine, indicates exposure to tobacco and other nicotine-containing products. This is an important part of wellness and screening programs, as smoking is tied to premature deaths and increases in lung cancer, heart disease, stroke, other cancers, osteoporosis, surgical complications, and numerous other health problems. In addition, tobacco use creates an economic burden in loss of productivity and absenteeism. Cotinine has a longer half-life than nicotine, which allows for a more accurate assessment of smoking status and better monitoring of smoking cessation progress. Because of its longer half-life and relative ease of detection, cotinine is the choice metabolite to assess smoking status. Testing for quantitative cotinine results with the PTS Detect cotinine system is quick and easy – let’s walk through the process.

Components Measures cotinine in capillary or venous whole blood Sample Size: 40 µL Measuring Range: 25 ng/mL – 200 ng/mL Measuring Time: 5 minutes after applying sample Available in 20-count kits The components of the PTS Detect™ cotinine system include the analyzer, test cartridges, shaker bodies, blood collectors and a package insert. It is important to remember that the components displayed above are pouched separately. We will discuss the testing process later in this training. We can use capillary or venous whole blood. Venous samples must be from a well-mixed green top lithium heparin tube. A 40 microliter blood sample is required for testing. The PTS Detect cotinine system has a measuring range of 25 to 200 nanograms per milliliter. Results are available in just 5 minutes after the sample is applied to the cartridge. The PTS Detect cotinine analyzer should be used only with the other components from its original kit. The analyzer will no longer run after the number of tests included in the original package have been used.

EXAMPLE: Expiration Date EXAMPLE: Room temperature storage time period Storage & Handling EXAMPLE: Expiration Date EXAMPLE: Room temperature storage time period REFRIGERATED (2 – 8ºC (36 – 46ºF)): Use until expiration date printed on box. ROOM TEMPERATURE (18-28ºC (64 – 82ºF)): Use until non-refrigerated time period printed on box, or up to expiration date, whichever comes first. IMPORTANT NOTES: If refrigerated, product should be removed from the refrigerator for one hour before testing to allow them to come to room temperature. Leave all components in their sealed pouches until use. Do not mix pouches and analyzers from different lots. Never use expired product. All components of the PTS Detect™ cotinine system should be stored refrigerated between 36 and 46 degrees Fahrenheit, or 2 and 8 degrees Celsius. Look for the expiration date label on your box – an example is shown on the left. When stored in the refrigerator, the PTS Detect cotinine system can be used until the expiration date printed on the box, analyzer and pouches. If desired, the PTS Detect cotinine system may be stored at room temperature prior to use for the time period printed on the box, or until the expiration date, whichever comes first. Look for the room temperature time period on your box – an example is shown on the right. Be sure to record the date that the kit was placed at room temperature. It is important to remove the product from the refrigerator for at least one hour prior to testing to allow the product to come to room temperature. Leave the pouch components in the pouch until the moment of use. Remember, do not interchange pouches and analyzers from different lots and never use expired product.

Testing Preparation The shaker and the blood collector are packaged in the shaker pouch. This pouch is labeled with a number 1 as it is used first in the testing process. The test cartridge is packaged in the other pouch. This pouch is labeled with a number 2 and should not be opened yet. To begin testing, gather the analyzer, the shaker pouch and the test cartridge pouch. Verify the lot number on each pouch matches the lot number on the back of the analyzer. You will find the lot number at the bottom of the pouches. Please see the example on your screen.

Sample Collection Use your own lancet device to draw blood. Gently touch blood drop to fill. Verify that both channels completely filled. Fully insert blood collector into shaker body. Twisting motion helps. Shake 6-8 times vigorously. This will mix the blood with the solution. Open the shaker pouch and remove the blood collector and the shaker body. DO NOT open the test cartridge pouch. Collect 40 microliters of blood either from a fingerstick or a venous lithium heparin sample using the blood collector. The blood collector has two channels and each channel must fill completely. The blood collector channels may need to be filled independently. Verify that both channels are completely filled. If collecting the sample from a fingerstick, please refer to the Fingerstick Technique Training, which is available at www.PTSDiagnostics.com/training. This separate training walks through how to conduct a good fingerstick. With the sample collected, quickly and firmly push and twist the blood collector into the shaker body, ensuring there is no gap or space between the shaker body and the blood collector. A twisting motion will help to ensure the blood collector is fully inserted. Vigorously shake 6 to 8 times to allow the blood to mix with the solution. Stand the shaker body on the table while you prepare for the next step.

Cartridge Preparation Caution: Do NOT open foil cartridge pouch until this step. Use within 2 minutes of opening. If foil pouch is damaged, do not use. Tear foil pouch open at the notches at the sides Retrieve the test cartridge pouch, labeled with a number 2. We can now open the cartridge pouch and remove the cartridge inside. REMEMBER - Until this point, do not open the cartridge pouch. The sample must be applied to the cartridge within 2 minutes after opening the cartridge pouch. Check that the codes on the test cartridge and the analyzer match, then insert the cartridge into the analyzer until you hear it click. Be sure that the analyzer is on a flat surface for the remainder of the test.

Analyzer Quality Check WAIT for SMPL to display Ready for shaker IMPORTANT NOTE: Wait for SMPL to appear on display before adding sample. This indicates the internal quality checks have been successfully completed. The analyzer will automatically begin some of its over 50 internal quality control checks. WAIT (ph. W-A-I-T) appears on the analyzer screen while these tests are being completed. Do not add the sample while WAIT (ph. W-A-I-T) appears on the screen. When the quality control checks have passed, the analyzer screen will show SMPL (ph. S-M-P-L) to indicate that it is now time to add the sample.

Applying the Sample Remove base from shaker body. Push firmly down on shaker, keeping the shaker vertical. Hold until “RUN” appears, then release. Do not move or handle analyzer until test is complete! To apply the sample to the cartridge, remove the base from the bottom of shaker, then align the circular bottom of the shaker body to the circular sample spot on the cartridge. Firmly press and release the shaker body on to the cartridge sample area and hold until “RUN” appears, then release. Also, ensure the shaker is kept upright in a vertical position relative to the cartridge during application. Do not apply the sample with the shaker body at an angle to the cartridge. When the sample has been detected, the analyzer will display RUN (ph. R-U-N). This means that the test is in process. At this point, the analyzer should not be touched or moved until the test is complete. IMPORTANT NOTE: It is very important to add the sample in one motion.

Reading Results After a 5-minute countdown, the analyzer will display three messages: QC test result (QCOK) Cotinine result (25 ng/mL – 200 ng/mL) Number of tests left (for example 01TL) The cotinine result and number of tests remaining will be displayed for at least 15 minutes or until the next test cartridge is inserted. Important Note: If QCOK, <25 or >200 is not displayed, an error code will be displayed in its place and no cotinine result will be shown. After a 5-minute countdown, the analyzer will display three messages: First, the analyzer will confirm that all quality control tests have completed successfully by displaying “QCOK” (ph. Q-C-O-K.) Next, the analyzer will display the cotinine result in nanograms per milliliter. Remember that quantitative cotinine results will display in the range from 25 ng/mL to 200 ng/mL. Results outside this range will either display less than 25 or greater than 200. Finally, the analyzer will display the number of tests left, for example, 01TL (ph. ZERO ONE T L) when there is 1 test left. The analyzer will continue to cycle through the display of these three messages for at least 15 minutes, or until a new cartridge is inserted in the analyzer. If “QCOK” (ph. Q-C-O-K.), less than 25 or greater than 200 is not displayed, an error code will be displayed in its place and no cotinine result will be shown. Refer to the package insert for help with error codes, or contact Customer Service.

Disposing of Test Cartridge Remove the cartridge and dispose of in accordance with your biohazard procedures. IMPORTANT NOTES: The analyzer is reusable. Do not discard. The analyzer will not perform any tests after the programmed number of tests have been run. (00TL indicates that all programmed tests have been used) After the cotinine result is recorded, remove the cartridge with gloved hands and dispose of the cartridge and shaker in accordance with your biohazard procedures. A new test may be started immediately following the same procedure. It is not necessary to wait for the analyzer to turn off prior to running the next test. A few tips to remember: The analyzer is reusable until all the cartridges in that kit are depleted or the number of tests left is zero, whichever comes first. 00TL (ph. ZERO ZERO T L) indicates that all programmed tests have been used. Note that if a cartridge is clicked in place in the analyzer and then removed before the sample is applied, it will consider it a test and count down the available tests left. Remember to only use the PTS Detect cotinine analyzer with the test supplies that came with it. Do not interchange between different kits. The analyzer will not perform any tests after the programmed number of tests have been run and can be disposed of as biohazardous electronic waste.