Obtaining blood samples for femme

1)Arrange for the blood samples to be taken on day 2, 3, or 4 of the woman’s menstrual period

2)OBTAIN WRITTEN, INFORMED CONSENT BEFORE YOU TAKE THE BLOOD SAMPLE

3)if other blood samples are being taken at the same time then please ensure the ‘order of draw’ is followed. for femme DRAW BLOOD INTO TWO SST (SERUM SEPERATOR TUBES) – THESE ARE USUALLY THE TUBES WITH YELLOW / GOLD CAPS, BUT CHECK THIS WITH your TRUST

4)GENTLY INVERT THE TUBES FIVE TIMES

5)If you do not have access to a centrifuge then place the sample into a royal mail safebox and post it to the Birmingham biobank who will process the sample. PLEASE ENCLOSE A COPY OF THE PATIENT’S CONSENT FORM AND A SAMPLE TRANSFER DOCUMENT WITH THE SAMPLES

6)if you do have access to a centrifuge then leave the tube to stand at room temperature for 20 – 30 minutes for a clot to form.Set the RPM on the centrifuge so it to sPINs AT 1,200g (this is not the same as the rpm setting, please check if you are uncertain)

7)if using a ‘swing out’ rotor spin the blood samples for 10 minutes. If the rotor has a fixed angle then spin the blood samples for 15 minutes

8)label two cryotubes with the patient’s intials, trial number and date sample was taken

9)Remove the tube from the centrifuge, and gently take the cap off

10)either decant or use the pasteur pipette to place the serum from one sst tube into one cryotube

11)cap the cryotubes and place them in the FEMME storage box in a -80°C freezer

12)Complete the sample log

BLOOD SAMPLE COLLECTION MANUAL

Staff Responsibilities

All personnel involved in taking blood samples or FEMME must sign the site Trial Signature & Delegation Logkept by the Principal Investigator. Personnel involved in obtaining blood samples as part of routine care do not need to sign this log.

Please ensure that the FEMME Trial office in Birmingham ()are informed of any changes in the contact details for the responsible staff. Should the responsible staff change, the TrialSignature & Delegation Log must be updated and forwarded to the FEMME Trial office.

1.Sample Collection Schedule

For FEMME, blood samples need to be taken at the following time points

1)Prior to the UAE or myomectomy procedure being performed

2)Six weeks after the procedure has been performed

3)Six months after the procedure has been performed

4)Twelve months after the procedure has been performed

In order to measure levels of some hormones we need to take the blood samples on either days 2, 3, or 4 of the woman’s menstrual cycle. By days 2, 3, or 4 we mean the second, third, or fourth day after flow has commenced or red blood is present, not the presence of spotting or old brown blood.

These blood samples will be used to measure the ovarian reserve.

2.Supplies

Each site will be supplied with blood sampling kits containing the following items:

  • Cryotubes and caps
  • Marker pen
  • Vacutainer® SST (Serum Separator tubes)
  • Disposable Pasteur Pipettes
  • Vacutainer venepuncture kit
  • Gloves
  • Royal Mail Safebox

When any of these items begin to run out then please call the FEMME Trials office on 0121 414 8335 or email us () for replacements.

The following equipment may be required but is not provided:

  • Centrifuge
  • -80°C Freezer (or -20°C if -80°C not available)

If the centrifuge is not available then once the sample has been taken place it into the secure Royal Mail Safebox and post it. The Royal Mail will deliver the sample to the Birmingham Biobank for processing and storage. If you do post the samples to us then please include a copy of the patient’s consent form and sample transfer form with the samples.

3.Obtaining the samples

Please consentthe patient before the blood samples are taken.

Samples for the FEMME study should be taken into TWO SST Vaccutainers® (these are usually the ones with the yellow / gold caps, but please check this before the blood is taken), i.e. a total blood sample of about 7 mls.

If other blood samples are being taken at the same time as those for FEMME then please observe the following ‘Order of Draw’ to avoid cross contamination of the tube contents:

1)Citrate tubes

2)No Additive

3)SST (gel) – FEMME SAMPLES

4)Heparin (gel)

5)Heparin

6)EDTA

7)Fluoride / Oxalate

Once the FEMME samples have been drawn into the SST tubesgently invert the tubes five times. If you do NOT have access to a centrifuge then label these tubes with the patient’s initials, trial number, date of birth and date the sample was taken.

If you DO have access to a centrifuge then leave at room temperature. Wait for 20 - 30 minutes for the clot to form then separatethe serum from the cells by spinning the tubes in a room temperature centrifuge at 1,2000g (rcf).

The rpm setting on the centrifuge necessary to reach 1200g will vary according to the length of its rotor but can be calculated from the following formula:

Where: r = distance in centimetres from the centre of the rotor to the position the material will be in when the centrifuge is spinning

Your answer can be checked by completing the ‘Calculate RPM’ boxes atbrokensymmetry.com/calc_rcf_rpm.htm

If the centrifuge is a ‘swing’ rotor spin these samples for ten minutes.

If the centrifuge has a fixed angle rotor then spin these samples for 15 minutes.

Once the samples have spun for the required amount of time remove the tubes from the centrifuge and gently remove the caps. Without disturbing the gel either decant, or use the pipette to draw off, the serum and transfer it to the cryotubes. These should be capped, labelled with the patients initials, trial number, dat of birth and date the sample was taken and placed in a -80°C freezer. Should this be unavailable then please place the cryotubes in a -20°C freezer.

Complete the sample collection information log (Appendix 1) for each sample taken. This information is necessary for us to ensure the integrity and quality of the samples.

Once the log is full send it to the FEMME Trials office either by fax it to 0121 415 9135, or email to. Please store the original log in the site file.

4.Sample Labelling

All cryotubes should be clearly labelled with the patient’s initials, date of birth, FEMME trial number and the date the sample was taken. The maker pen supplied (Pentel N50) is safe to use on samples that will be stored for a long time. If you use another labelling method then please ensure that it is unaffected by long term storage at -80°C.

5.Sample Storage

Samples should be stored in a freezer at -80°C. Once a suitable number of samples have been accrued then place these samples into the Royal Mail Safebox and put them in the post for delivery to Birmingham’s Biobank. If you start to run out of Safeboxes then just email r some more. When returning samples to us, please include a copy of the patient’s consent form and sample transfer log in the Royal Mail Safebox.

5.1.Storage Conditions

All serum samples should be stored in a -80°C freezer. If this is unavailable then the samples may be stored in a-20°C freezer.

5.2.Temperature Deviation

During storage the freezer temperature must be monitored regularly. Ideally this should be a continuing electronic monitoring system, however an established system of recording min / max thermometer readings once a day is acceptable. Temperature logs should be maintained at all times and should be available for inspection by the monitoring team on request.

Should the temperature in the freezer ever be greater than -5°C please inform the FEMME Trial office on 0121 414 8335 or by email at ld the temperature exceed -5°C this must be recorded in the Temperature log.

6.Transportation of Samplesto a central biobank

Once a number of samples have accrued then please place the samples into the supplied Royal Mail Safebox and put them in the post for delivery to Birmingham’s Biobank. If you start to run out of Safeboxes then just r some more.

.

7.Monitoring

The FEMMETrial will be monitored by the FEMME Trial Coordinator, representatives of the sponsors (Oxford University) andthe regulatory authorities.

8. Contacts

Should you have any queries or require further information, please contact:

William McKinnon (Trial Coordinator)

Birmingham Clinical Trials Unit

Robert Aitken Institute

University of Birmingham

Edgbaston

Birmingham

B15 2TT

: 0121 414 8335

FAX: 0121 415 9136

APPENDIX1- SAMPLE COLLECTION INFORMATION LOG

SAMPLE COLLECTION INFORMATION LOG

HOSPITAL:
Patient Initials / Date of Birth / Date of Sample / Day of period
(2, 3 or 4) / Date sample spun and put in Freezer / Date sent to Biobank / Your name
(CAPITAL LETTERS) / Signature / Comments
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
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DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY
DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY / DD/MMM/ YYYY