Cleveland Family Study

3.5.2 Venipuncture Procedure

ALWAYS WEAR LATEX GLOVES AND LAB COAT.

1. Arrange draw tubes in order of draw on the table top or in the tube rack within easy reach. Assemble butterfly apparatus and Vacutainer holders, gauze, and alcohol prep prior to tourniquet application.
2. Apply tourniquet.
3. Examine participant's arms for the best site for venipuncture. Release tourniquet
4. Cleanse venipuncture site by wiping with alcohol prep pad in a circular motion from center to periphery. Allow area to dry.
5. Reapply tourniquet and start timer.

   Note: If possible, it is best to release the tourniquet as soon as possible after flow has been established.

   • Tightened tourniquet should be on no longer than 2 minutes recommended or loosen tourniquet, then reapply if necessary. In our experience, however, especially with sick and/or elderly subjects, this may result in flow stopping, and the trauma of a second venipuncture. Therefore, this is a judgment call based upon the phlebotomist’s experience and skill.
6. Grasp the participant's arm firmly, using your thumb to draw the skin taut. This anchors the vein. The thumb should be 1 or 2 inches below the venipuncture site.
7. With the needle bevel upward, enter the vein in a smooth continuous motion.
8. Make sure the participant's arm is in a flat or downward position while maintaining the tube below the site when the needle is in the vein. It may be helpful to have the participant make a fist with the opposite hand and place it under the elbow for support.
9. Grasp the flange of the Vacutainer holder and gently push the tube forward until the butt end of the needle punctures the stopper, exposing the full lumen of the needle.

   Note: Attention should be paid to minimizing turbulence whenever possible.

   • Small steps, such as slanting the needle in the Vacutainer to have the blood run down the side of the tube instead of shooting all the way to the bottom, may result in significant improvement.
10. Note the blood flow into the first collection tube.
    • If blood is flowing freely, the butterfly needle can be taped to the participant's arm for the duration of the draw.
    • If the flow rate is very slow, the needle may not be positioned correctly. Try moving the needle slightly without causing discomfort to the participant.
11. Keep a constant, slight forward pressure (in the direction of the needle) on the end of the tube. This prevents release of the shutoff valve and stopping of blood flow. Do not vary pressure nor reintroduce pressure after completion of the draw.
12. Fill each Vacutainer tube as completely as possible (e.g., until the vacuum is exhausted and blood flow ceases). If a Vacutainer tube fills only partially, remove the tube and attach another without removing the needle from vein.
13. When the blood flow ceases, remove the tube from the Vacutainer holder. The shutoff valve re-covers the point, stopping blood flow until the next tube is inserted (if necessary).
14. Place the EDTA, Citrate and SCAT-1 tubes on tube mixer for a minimum of 30 seconds. The EDTA, Citrate and SCAT-1 tubes may also be mixed manually by gently inverting the tubes for at least 30 seconds. Do not mix the serum tubes.
15. Release tourniquet, if still applied.
16. To remove the needle, lightly place clean gauze over venipuncture site. DO NOT have the patient bend his/her arm as this can cause a hematoma.
17. Remove the needle quickly and immediately apply pressure to the site with a gauze pad.
18. Have the participant hold the gauze pad firmly for one to two minutes to prevent a hematoma.
19. Discard needle into puncture-proof sharps container.
20. Record on Phlebotomy form duration tourniquet was applied and length of venipuncture.
21. The EDTA, Citrate and SCAT-1 tubes are placed on wet ice. The serum tubes are held at room temperature.
22. Clean up the venipuncture area (if necessary).
23. Dispose of needle and tubing in the appropriate biohazard needle sharps containers.
24. Complete the Phlebotomy Form
25. Bring the filled blood collection tubes to the processing area, keeping the EDTA, Citrate and SCAT-1tubes on ice and the serum tubes at room temperature

Since there are multiple draws involved in this protocol, care must be taken in selecting the second and third draws sites. This is most important for the second draw at 8:00 AM where care should be taken to avoid the previous draw site. Preferably, the arm not used for the first PM blood draw should be chosen for the second draw. If this is not feasible (only one “good” vein or both arms were stuck to get the first draw), the preferred site would be above the first draw site.

3.5.2.1 Oral Glucose Procedure

Oral glucose will be administered after fasting, post-sleep venipuncture is performed.

1. After final awakening and prior to rising and eating in the morning, oral glucose will be administered by study nurse (75 grams or up to 1.75 g/kg in children).
2. Have patient lie down after drinking.
3. Approximately 2-hours after administration, draw 5 mL blood in serum tube. Use a 21-guage butterfly needle with 12 inches of plastic tubing between the venipuncture site and the blood collection tubes. Use the same procedure as outlined above.