Blood and Urine Processing for PNG

Check freezer and refrigerator space availability and temperatures. Freezer space
should have been previously allotted for use with a phone call or another contact ahead
of time.

Take samples to the University of Papua New Guinea (UPNG), School of Medicine and
Health Sciences, Division of Basic Medical Sciences.

Place digital thermometers into the freezer and refrigerator, so that you may record the
temperatures onto all shipping lists.

Unpack specimens and note comments or problems on Sample Receipt List (e.g.,
sample leaked, not labeled correctly, or samples too warm).

Store urine samples in a -20°C freezer and purple tops in 4°C refrigerator while
processing red tops. Purple tops may be discarded into a biohazard container after the
zinc protoporphyrin has been recorded and the dried blood spots have been pipetted.

Because of Vitamin A’s sensitivity to light, do not pipette the dried blood spots near
sunlight from a window. Fluorescent lighting is okay.

Centrifuge red tops according to manufacturer’s instructions for tube size to separate the
serum from the cells.
Red Top Processing:
Preparations
Organize red top tubes, racks, disposable pipettes, cryovials and caps, Sample Receipt
List, shipping list, boxes, study labels and box labels to facilitate good work flow.

Thirteen (10X10) gridded boxes for serum cryovials should be prelabeled on the side of
the lid and then handwritten/prelabeled on the side of the box bottom. 8 boxes will be
shipped to CDC and 5 boxes will stay with Victor.

Boxes need to be marked at the top left corner with a heavy marker. This will indicate
specimen placement in box (i.e., working left to right, back to front in box.)

Placing a label on the front side of the box helps for ID of box for freezer storage.
Processing
Centrifuge red top tubes according to the manufacturer’s instructions.

After red top tubes are centrifuged, organize the tubes in a rack corresponding to the
order of the labels.
Take the 5 labels identified as Serum belonging to the first patient (according to the label
ID on the red top tube) and prelabel the serum cryovials. Work with one participant ID
at a time and label the shipping list and the Sample Receipt list accurately. Note
any comments.

If there is an “unable to draw”, make sure that the shipment list and the Sample Receipt
list indicate this.

Working with one specimen at a time always check label ID’s on all tubes and double
check on transfer from clot tube to cryovials.

Pipette at least ½ mL serum into each of the 5 cryovials utilizing all available serum. 3
cryovials containing ½ mL serum each are necessary for testing at CDC, and the
remaining two will stay at UPNG for use in establishing the Division of Basic Medical
Sciences laboratory’s vitamin A method. NEVER overfill beyond the 1.8 ml mark (This
is to allow for freezer expansion).

Work quickly, and secure caps tightly after each transfer.

Discard each pipette into a biohazard bag and each blood tube into a sharps container.

Place 3 cryovials into one of 8 boxes (to be shipped to CDC) and the other two vials into
one of 5 boxes (to stay with VT).

After all red top serums have been transferred to labeled cryovials, securely capped, and
appropriately boxed, place the boxes into the -20°C freezer (in the designated area and
leave a notation on freezer door indicating the 2005 PNG Validation Study) until ready to
package the 8 boxes for shipment to CDC.

Make 2 copies of the shipping list. The original list will stay with the samples (give to VT
), the two copies will go to BB..

The sera will be shipped back to CDC via World Courier by BB during the week of May
23rd, 2005.
Purple top Processing:

Retrieve purple top tubes from the 4°C refrigerator.

Organize 25 microliter pipette and tips, DBS cards, sharps container,
hematofluorometer, cover slips, controls, ZP quality control log, ZP results form,
shipping list for DBS cards, and labels to facilitate good work flow.

Using the tube racks, organize the purple top samples into sequential order and mark on
the Sample Receipt List.

Place 4 DBS labels (of the same ID # as the first purple top tube) on the first DBS card.

Follow the protocol for zinc protoporphyrin (ZP) testing on the first tube (with the same
ID # as the DBS labels on the card), being sure to invert the tube before using to ensure
a homogeneous sample.

Once the ZP testing has been done for that sample and the result has been recorded,
invert the capped tube to mix once more and pipette the blood onto a DBS card, filling
all 8 circles and then the spaces beside the circles, if possible. Place the DBS card
horizontally into the DBS box and allow to dry. Discard the used pipette tip into the
biohazard bag and the purple top tube into the sharps container.

Place 4 labels corresponding to the ID # on the next purple top tube on another DBS
card. Perform the ZP testing and DBS pipetting using this sample. Repeat this process
with the rest of the purple top tubes.

Once the DBS cards are dry, follow the “DBS Card Preparation for PNG” protocol for
packing and storage.

Testing for ZP is stable for up to 7 days if stored in a refrigerator. Do Not Freeze.
Final Tasks:
 Before team workers leave the processing lab, make sure the lab area is as it was on
arrival.
o Remove all items that you brought in.
o Discard all trash into appropriate containers.
o Don’t forget your digital thermometers.
o Xerox more blank copies of Shipping Lists.
o Turn off the Xerox machine.
o Place used cold packs in -20°C freezer.
Refer to the shipment procedure when ready to package and ship the sera and DBS cards to
CDC.