Standard Operating ProceduresCollection of Blood Specimen: Dried Blood

Spot

Purpose: Clients who receive a 'recent' test result on the POC test will be asked to provide a specimen for viral load testing at the laboratory, which will be used to confirm the recency test result. For the [name of country] HIV recency testing, capillary blood will be collected via finger prick for the preparation of dried blood spot (DBS) specimens. This standard operating procedure describes procedures for collecting DBS specimens.

Staff Responsible: Nurse or HTS Provider (to be assigned in each facility)

Materials Needed: Forms:

Laboratory Requisition Form (1) Recent HIV Surveillance Register* (1) - [dependent on country context]

Other materials: Whatman 903 DBS card (1), Becton-Dickinson, blue lancet (1), gauze square (1), alcohol swab (1), plaster (1), desiccant packs (3), sealable plastic bag (1), capillary collection tubes (2); disposable gloves (2); Participant ID Label * (2 - remove if your country is not using barcode label stickers); ballpoint pen (1); sharps container (1); paperclip (1); and drying rack (1)

Procedures: 1. Organize all supplies needed for preparation of DBS. 2. Using a ballpoint pen, carefully affix a Participant ID Label or write

the Participant's ID Number on the Whatman 903 DBS card. Write the date and time of blood collection.

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If using a pre-printed Participant ID Label, take care to place the label below the first black line, so it does not touch the blood spots.

3. Wash your hands thoroughly and put on a clean pair of disposable gloves.

4. Position the participant’s hand palm-side up. Choose the finger that is least callous (usually the third or fourth finger). Do not use the same finger that was used for the HIV test.

5. Massage the participant’s finger by gently squeezing from the base of the finger to the fingertip 5-6 times to increase blood flow.

6. Use an alcohol swab to sterilize the finger and allow to air dry for 30 seconds.

7. Hold the finger and firmly place a new sterile lancet off-center on the fingertip.

8. Firmly press the lancet to puncture the fingertip.9. Use a clean gauze square to wipe away the first drop of blood from the

puncture site. 10. Collect drops of blood into the capillary collection tubes by gently

massaging the finger. 11. Have patient hold a gauze pad or cotton ball to the puncture site

until the bleeding stops. Once preparation of DBS card complete, return and cover the puncture site with a plaster.

12. While waiting for bleeding to stop, prepare DBS card: Lightly touch the DBS card to the end of the capillary collection

tubes and allow the blood to flow onto the filter paper. Allow blood to soak through the card until the circle is completely

full. Repeat until all five circles on the DBS card are filled. The size of

the blood spot should completely fill the sample circle (equivalent to one large drop of blood). Avoid touching or smearing the blood spots.

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13. Place card on drying rack. Allow the specimen to fully air dry horizontally (at least 4 hours) at room temperature.

14. Remove and dispose gloves and other biohazardous material. Dispose lancet in sharps container provided.

14. Complete Sections 1 and 2 of the Laboratory Requisition Form using a ballpoint pen.

Write the facility name, facility code, and date of the participant’s HIV diagnosis.

Write the Client Identification Number (ART/HTS/ANC number), specimen type: check one – venous blood or DBS. Affix Participant ID Label to form or write Participant’s ID Number on space provided.

Write the date and time of specimen collection and the name of the staff collecting the specimen.

Leave Sections 3-6 blank to be completed later.15. If using the Recent HIV Surveillance Register, complete column

indicating whether a specimen was prepared for additional testing.

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Standard Operating ProceduresCollection of Blood Specimen: Venous

BloodPurpose: Clients who receive a 'recent' test result on the POC test will be asked to provide a specimen for viral load testing at the laboratory, which will be used to confirm the recency test result. For the [name of country] HIV recency testing, approximately 5ml of venous blood will be collected from participants in ethylene-diamine-tetra-acetic acid (EDTA) vacuum tubes (vacutainer tubes). This standard operating procedure describes procedures for collecting venous blood specimens.

Staff Responsible: Nurse (to be assigned in each facility)

Materials Needed: Forms:

Laboratory Requisition Form (1) Recent HIV Surveillance Register* (1) - [dependent on country context]

Other materials: Tourniquet (1), alcohol swab (1), gauze square (1), plaster (1), adult butterfly needle set (1), vacutainer single-use holder (1), ethylene-diamine-tetra-acetic acid (EDTA) vacuum tube - purple top (1); Participant ID Label* (2 - remove if your country is not using barcode label stickers), disposable gloves (2); ballpoint pen (1); ultra-fine permanent marker (1); sharps container (1)

Procedures: 1. Organize all supplies needed for venous blood collection. 2. Wash your hands thoroughly and put on a clean pair of disposable

gloves. 3. Assemble needle and vacuum tube holder. Attach the end of the

winged infusion set to the end of the vacuum tube and insert the

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collection tube into the holder until the tube reaches the needle. Remove the plastic sleeve from the end of the butterfly.

4. Put a tourniquet 7 to 10cm above the venipuncture site (the bend of the elbow).

5. Ask the participant to make a fist with her hand so that the veins become more prominent.

6. After palpating veins, use an alcohol swab to sterilize the puncture facility and allow the area to air dry for 30 seconds.

7. Press the butterfly tips together and remove the protective sheath from the needle.

8. Position the needle with the bevel facing up. Use your thumb to tighten the skin around the venipuncture site.

9. Enter the vein at a 15-to-30-degree angle. Spread open the butterfly wings and securely rest them flat against the skin.

10. Slowly push the tube forward to start blood flow. Allow the tube to fill to obtain 5 mL of whole blood. Release the tourniquet.

11. Have the participant release the fist and relax the tourniquet.12. Remove the tube and gently invert it 8-10 times before laying it

down. This will prevent the blood specimen from clotting. 13. Place a clean gauze square on the venipuncture facility and

slowly withdraw the needle. Immediately move the gauze over the puncture facility and apply pressure to stop bleeding and avoid formation of hematoma.

14. Remove the butterfly needle from the vacuum tube holder and discard immediately in appropriate sharps container. Continue applying pressure with gauze to the vein until bleeding stops.

15. Check to ensure there is no hematoma formation and then apply plaster to puncture facility.

15. Use a permanent marker to note the date of specimen collection. Affix a Participant ID Label to the EDTA tube if using pre-printed barcode labels; otherwise, write the Participant's ID Number in the space provided.

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16. Remove and dispose gloves and other biohazardous material. Dispose of butterfly needles in sharps container provided.

17. Complete Sections 1 and 2 of the Laboratory Requisition Form using a ballpoint pen.

Write the facility name, facility code, and date of the participant’s HIV diagnosis.

Write the Client Identification Number (HTS/ART/ANC number), specimen type: check one – venous blood or DBS. Affix a Participant Identification Label to form or write the Participant’s ID Number in space provided.

Write the date and time of specimen collection and the name of the staff collecting the specimen.

Leave Sections 3-6 blank to be completed later.18. If using the Recent HIV Surveillance Register, complete column

indicating whether a specimen was prepared for additional testing.

16.

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APPENDIX A

Supply Checklist for Dried Blood Spot Collection, Handling, and Storage

BD, blue lancet

(1)

Alcohol swab

(1)

Gauze square (1) Capillary tubes (2)

Whatman 903 DBS card (1)

Plaster (1)

Desiccant packets (3)

Plastic bag (1)

Disposable gloves (2)

Sharps container (1)

Drying rack (1) Specimen label (1)

Envelope (1) Paper Clip (1) Lab Requisition Form

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APPENDIX B

Supply Checklist for Venous Blood Collection, Handling, and Storage

Adult butterfly needle set (1)

Vacutainer single-use holder (1)

EDTA, purple top vacuum tube (1)

Tourniquet (1)

Alcohol swab (1) Gauze square (1) Plaster (1) Disposable gloves (2)

Sharps container (1)

Specimen label (1)

Cardboard Storage Box (1)

Ice Packs (2)

Ultrafine Permanent Marker (1)

Cooler(1) Lab Requisition Form

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