Attachment_1_-Human_Sera_Collection_Guidelines.pdf

LP-093, R-2D (M. Levine) Distribution Copy Effective: April 24, 2013 Page 1 of 1

Human Sera Collection Guidelines for Flu Serology Immunology and Pathogenesis Branch, Influenza Division

Collection Details

1. We suggest collecting the following volumes of blood:

Category Age of blood donor Volume of blood to collect* Pediatric Less than 3 years old 5.0 ml Children 3 through 11 years old 5-10.0 ml Teens, adults, and elderly 12 years and older ≥10.0 ml

*For teens, adults and elderly persons we request that at least 10 ml of blood be drawn. Sera will be tested in multiple assays and potentially with multiple viruses.

In general, while the above volumes are preferred, smaller volumes will be accepted, if the situation does not allow for the collection of these amounts, e.g. for infants and young children. The minimum accepted volumes of blood to collect are 5ml from adults and older children and 1.5 ml from younger children.

2. Use tubes designated for the collection of serum, not plasma. We suggest the use of vacutainer tubes.

The following vacutainer tubes are acceptable:

a. glass red top vacutainer tubes

b. plastic red top vacutainer tubes with clot activator

c. plastic gold top serum separator tubes (SST) with clot activator and gel

3. For testing of serum samples in the microneutralization assay, it is very important to minimize hemolysis.

Hemolyzed serum often has a negative impact on the cells in the microneutralization assay. We recommend the use of a butterfly needle connected to a vacutainer tube as the best collection method to minimize hemolysis and to reduce the risk of needle stick injury. The butterfly needle is a safe one-way system with blood being delivered into sealed vacutainer with a consistent pressure flow. The only source of possible hemolysis with this system is if the needle in the vein scrapes against vein wall and blood cells break up as they enter the needle bore. As an alternative choice, the use of a needle that connects directly to the vacutainer tube is acceptable.

Storage Recommendations

Collected blood should be stored at 4°C immediately. This can be done by placing the sample on ice, in a 4°C refrigerator, or in a cooler with cold packs. Blood may be stored at 4°C for up to 18 hours. Sometime between 1 hour and 18 hours after collection, the blood collection tube must be centrifuged to separate the clotted blood from the serum and the serum removed to a clean tube. The clotted blood may be discarded. The serum must be aliquoted into labeled tubes. After aliquoting, the serum samples must be immediately stored at -20°C or colder.

Shipping Guidelines – Send questions and tracking info to: MLevine@cdc.gov and HTatum@cdc.gov. Send serum samples to:

CDC - Flu Serology Attn: Ms. Heather Tatum Bldg 23, Room 8451 1600 Clifton Road, NE Atlanta, GA 30333 Phone: 404-639-4130

All shipments should be via overnight carrier, to ensure delivery within 24 hours of shipment. All serum shipments should be packed with sufficient dry ice to ensure that samples remain frozen for a minimum of 48 hours after shipment.