1. Collection, Storage and Transportation of Dried Blood Spots for Infant Diagnosis

2. Learning Objectives Collect Dried Blood Spots (DBS) for HIV testing
Package and store DBS so as to maintain specimen integrity
Distinguish between valid and invalid DBS

3. Step 1 Equipment DBS sample collection card DBS Lab requisition Form
Sterile lancet
Sterile alcohol prep
Sterile gauze/cotton swab
Gloves
Drying rack
Low Gas Permeable zip-lock bag
Desiccant
Humidity indicator card
You should assemble all the equipment you need before hand.

4. Step 2 Correctly complete all the information on the requisition form
Keep copy for your records

5. Step 3 Wash hands and dry thoroughly Put on gloves
Confirm identity of the infant and write the baby’s name, date of the test and the unique identifier on the filter paper
Do no not contaminate by touching or allowing spillage on the filter paper
Jane Doe
May 12th 2006
Do not allow water, formulae, powder from gloves, antiseptic solutions to come into contact with filter paper before or after collection of sample

6. Step 4 Choosing the site Small infants (<9kg)
Puncture the heel.
Do not puncture the fingers; there is risk of hitting the bone.
Larger infants (> 9kg)
Puncture the heel; if callous is visible, you may use the lateral aspect of the big toe.
Do not stick the fingers or small toes; there is risk of hitting the bone.
Fingers are safe in children > 2 years of age
This is also true when doing an HIV rapid test. Do not stick fingers or small toes of small children. Fingers are safe around age 2)

7. Step 5
Do not puncture here
Safe
Draw an imaginary line from midpoint of the big toe to the heel and one from between the 4th and 5th toe to the heel.
Shaded area indicates safe areas for puncture site
Do not puncture the back of the heel or Achilles tendon or the medial aspect of the big toe
Draw an imaginary line from midpoint of the big toe to the heel and one form the between the 4th and 5th toe to the heel.
safe
safe
Do not puncture here

8. Step 6 Position baby with foot down
This will help the blood flow more easily
Warm site with soft cloth moistened with warm water up to 410c for 3 to 5 minutes

9. Step 7 Clean site with alcohol prep Allow to air dry for 30 seconds
Failure to allow alcohol to dry may dilute the specimen
Failure to allow alcohol to dry may dilute the specimen

10. Step 8
Puncture the toe to a depth less than 2 mm using a sterile lancet

11. Step 9 Wipe away first blood with sterile gauze pad
The initial drop contains tissue fluid that may dilute the specimen
Allow another large blood drop to form

12. Step 10 Lightly touch the filter paper to the large drop of blood
Allow to soak through and completely fill the circle
To enhance flow very gently apply intermittent pressure to area above the puncture site.
Excessive milking or squeezing the puncture may cause hemolysis and mix tissue fluid with specimen affecting the results

13. Step 11
Apply blood to one side of the filter paper only, the side with the printing
Do not layer successive drops of blood or apply blood more than once to the same collection circle
Avoid touching or smearing the spots

14. Step 12
Fill remaining circles in the same manner with successive drops of blood
If blood flow is diminished repeat steps 10 and 11

15. Step 13
Inspect card to ensure you have collected enough blood, and the specimen is valid
Jane Doe
May 12th 2006
Do not allow water, formulae, powder from gloves antiseptic solutions to come into contact with filter care before or after collection of sample
Valid Specimen

16. Step 14
Place filter paper on a horizontal clean dry surface to AIR DRY for at least 3 hours
Keep away from direct sunlight, dust, and bugs
Do not allow blood spots to come into contact with any surface or each other.
Do not stack , heat or expose to excessive sunlight during drying. Allow to dry completely before placing in the Ziploc bag. Insufficient drying will affect the results.

17. Step 15
Place dried DBS cards between sheets of glassine paper so that the DBS cards are not touching each other
Fold ends of the glassine paper

18. Step 16 Place dried filter paper into Ziploc bag Add desiccant package
Minimum of 2 desiccant packages per specimen
It is safe to add at least 10 silica desiccant packages per bag.

19. Step 17 Add humidity indicator card Remove air and seal bag
Keep packaged DBS (in sealable plastic bags) refrigerated until transported to reference laboratory

20. Step 18 Transportation Place bag into envelope
Add lab requisition slips and specimen delivery check list
Clearly label outside of the envelope
Transport to central laboratory for processing

21. Step 19
During transportation do not leave in vehicle, as sun and heat will deteriorate DBS

22. Invalid Specimen Specimen quantity insufficient for testing
Removing filter paper before blood has completely filled circle or before blood has soaked through to second side.
Applying blood to filter paper with a capillary tube
Touching the filter paper before or after specimen collection with a gloved or ungloved hands, hand lotion etc.
Allowing the filter paper to come into contact with gloved or ungloved hands or stances such as hand lotion or powder, either before or after blood specimen collection.

23. Invalid Specimen Specimen appears scratched or abraded
Applying blood with a capillary tube or other device

24. Invalid Specimen Specimen not dry before mailing
Mailing specimen before drying for a minimum of 3 hours

25. Invalid Specimen Specimen exhibits serum rings
Not wiping alcohol from puncture site
Allowing filter paper to come into contact with alcohol , hand lotion etc.
Squeezing area surrounding puncture site excessively
Drying specimen improperly
Applying blood to filter paper with capillary tube

26. Specimen appears diluted, discolored
Invalid Specimen
Specimen appears diluted, discolored
or contaminated
Squeezing or milking of area surrounding puncture site
Allowing filter paper to come into contact with gloved or ungloved hands or substances such as alcohol, formula, antiseptic, water, powder etc. before or after blood specimen collection
Exposing blood spots to direct heat

27. Invalid Specimen
No blood
Failure to obtain specimen

28. Specimen appears clotted or layered
Invalid Specimen
Specimen appears clotted or layered
Touching the same circle on filter paper to blood several times
Filling circle on both sides.

29. Specimen appears super saturated
Invalid Specimen
Specimen appears super saturated
Applying excess blood to filter paper, usually with a device
Applying blood to both sides of filter paper.