Nov 21, 2022

Public workspaceBlood Draw

DOI
dx.doi.org/10.17504/protocols.io.261ge4w5dv47/v1
  • Clemens Scherzer1,2,
  • Bradley Hyman3,2,
  • Charles Jennings1,2
  • 1Brigham and Women's Hospital;
  • 2Harvard Medical School;
  • 3Massachusetts General Hospital
Daniel El Kodsi
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DOI: dx.doi.org/10.17504/protocols.io.261ge4w5dv47/v1
Protocol Citation: Clemens Scherzer, Bradley Hyman, Charles Jennings 2022. Blood Draw. protocols.io https://dx.doi.org/10.17504/protocols.io.261ge4w5dv47/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 18, 2021
Last Modified: May 31, 2024
Protocol Integer ID: 47406
Keywords: blood, draw, ASAPCRN
Abstract
This protocol explains the Standard Operating Protocol for Blood Drawing.
Guidelines
PROTOCOLS NO LONGER IN USE

SERUM
MATERIALS: One 4 ml yellow top SST Gel and Clot Activator SERUM tube (BD Franklin Lakes, NJ., Ref# 367977) PROTOCOL: After centrifugation, pipette 500 uL of the liquid phase above the formed clot into each of the four tubes labeled “SE-01, SE-02, etc.” Avoid contaminating the sample by preventing the pipette tip from touching the wax that the serum sits upon. Tipping of the tube can ease the access to the remaining serum volume that sits on top of the wax layer. The remainder of the tube can be discarded.


FREEZER STORAGE

Freezers are divided into 4 shelves, with 6 racks per shelf, and 24 boxes that can be held in each shelf. In total, 576 boxes, approximately 2,160 sample sets, can be stored in one -80°C freezer. The first three shelves are designated by visit number: Shelves A1-6 (top shelf) house samples from enrollment visits, shelves B1-6 (2nd shelf) house samples from the 1st year follow-up, and shelves C1-6 (3rd shelf) house samples from the 2nd year follow-up. Shelves D1-6 contain packed red blood cell tubes (PRBC), DNA, and RNA, extracted from blood as described in the protocols above. CSF is designated between two freezers in selected racks. Freezer storage and transactions of samples are recorded in the Freezerworks Inventory software.
Materials
MATERIALS:
  1. “Butterfly” 21- or 23-gauge Vacutainer needle with adapter for tube holder
  2. Tube holder (Smiths Ref# 4142)
  3. Individually packaged 70% isopropyl alcohol wipes
  4. Latex-Free Bandaids or Tape
  5. Latex-Free Gauze Cover Sponges
  6. Biohazard Sample Bags (Fisher Ref# 18-028321)
  7. One Latex-Free Tourniquet
  8. Latex-Free Gloves
  9. Sharps Disposal Container. An OSHA acceptable, puncture proof container marked "Biohazardous"

SPECIMEN TUBES:
  1. Two 2.5 cc PAXgene™ Blood RNA Tubes (VWR Ref# 77776-026)
  2. One 10 ml BD purple top EDTA-BD Vacutainer tube (BD Franklin Lakes, NJ., Ref# 366643)
  3. One 8.5 ml black and blue CPT tube- (BD362761)
  4. Two 6 ml purple top EDTA-BD Vacutainer tubes (BD Franklin Lakes, NJ., Ref# 367863)
  5. One 2 ml purple top EDTA-BD Vacutainer tube (BD Franklin Lakes, NJ., Ref# 367841)
  6. One 2.7 ml blue top EDTA-BD Vacutainer tube (BD Franklin Lakes, NJ., Ref# 363083)
  7. One 8.5 ml BD ACD Solution A Vacutainer tube (BD Franklin Lakes, NJ., Ref# 364606)

***Note: Please see appendix in guidelines for information regarding SERUM tube and processing. SERUM collection was discontinued 5/1/2019
Safety warnings
Please refer to Safety Data Sheets (SDS) for health and environmental hazards. Gain all required consent and experimental approvals before beginning any procedures.
Blood Draw
Blood Draw
10s
10s
Store all tubes at TemperatureRoom temperature (18ºC to 25ºC) before use.

Label all tubes with the subject ID# prior to the blood draw.
The PAXgene™ Blood RNA Tubes should be the first tubes drawn in the phlebotomy procedure. Collect all other tubes after collecting the PAXgene™ Blood RNA Tubes in the following order: 10 ml EDTA, one 8.5 ml black and blue 8.5 ml CPT tube, two 6 ml EDTA, 2 ml EDTA, 2.7 ml EDTA and 2.7 ml blue top (only if interested in doing an LP). In special cases, one 8.5 ml Solution A will be substituted for one 6ml purple top EDTA-BD Vacutainer tube (only taken once per subject if applicable). When applicable, a 2.7 ml EDTA tube is taken last.
Prior to blood draw, ask donor three questions:
  1. Are you allergic to latex?
  2. Do you take blood thinners?
  3. Do you have a history of fainting during blood draws?
If donor answers yes for any question, take appropriate precautions. Ask donor to sit, place pillow (covered with blue pad) beneath arm. Examine donor’s arm for optimal puncture site, then apply tourniquet. Using gloves, clean site with alcohol wipes.
Using a blood collection set and a holder, collect blood into the first PAXgene™ Blood RNA Tube using your institution’s recommended procedure for standard venipuncture technique.

The following techniques shall be used to prevent possible backflow:
Place donor’s arm in straight, downward position.
Hold tube in a vertical position, below the donor’s arm during blood collection.
Release tourniquet as soon as blood starts to flow into tube.
Make sure tube additives do not touch stopper or end of the needle during venipuncture.
Allow at least Duration00:00:10 for a complete blood draw to take place in each tube. Ensure that the blood has stopped flowing into the tube before removing the tube from the holder.

10s
Immediately after blood collection, gently invert/mix (180 degree turns) the tubes 8 – 10 times.
Mix
REPEAT STEPS 4 to 6 for each of the tubes to be collected per subject. Go togo to step #4 .

Remove needle in quick motion and dispose in Sharps Disposal Container. Immediately press gauze to puncture site, ask donor to apply pressure and raise arm (keeping it straight) until blood flow stops. Bandage with Latex-Free Bandaids or gauze and tape.
Record time/date of draw on specimen collection form. Attempt to keep all tubes upright.
Place the 8.5 ml Solution A tube in a separate biohazard bag along with the requisition form including gender, date drawn, and date sent information. Ideally within 4 days of the draw, it should be delivered by cooler to the lab that forms the cell line. It should be kept at TemperatureRoom temperature (18ºC to 25ºC) until then.

The 2 ml purple top CBC will be sent in a separate biohazard bag the same day of the draw to the core lab for processing immediately along with the proper requisition forms and labeling. Reticulocytes are requested at the enrollment visit only.
The rest of the tubes are placed in a biohazard bag (double bag if necessary) and are brought to the lab by cooler within 3 hours of the draw for processing and storage.
2.7 ml blue top tube is placed on ice in a specimen cup and sent to the core lab for immediate PTT/PT/INR processing with the proper requisition forms and labeling.