Preparation of QC and Training Panels International Laboratory Branch Division of Global HIV/AIDS & Tuberculosis Centers for Disease Control and Prevention

Outline Describe the considerations for preparation and characterization of a specimen bank Review the step-by-step process Explain the components within the QC and Training panels Summarize the verification process for the QC and training panel Define conditions for usage of the panels

Prior to the TOt

General Considerations for a Specimen Bank 4 General Considerations for a Specimen Bank Obtain rejected whole blood units from the blood bank The blood specimen must be re-tested using the rapid test algorithm (National HIV RDT algorithm) Process confirmed HIV positive and few negative blood units into plasma Centrifuging the blood at 3000 rpm for 10 minutes Test HIV positive specimens with RTRI to determine recent or LT status Continue collecting, re-testing, and characterizing the positive blood specimens to expand the collection for future HIV research A small/sizable number of negatives should be included in the collection Also mention for the specimen bank that negatives should also be stored in the collection, however the laboratory only needs to store several tubes.

Characterization of HIV-1 Positive Samples 5 Characterization of HIV-1 Positive Samples Characterize HIV positive specimens using rapid test for recent infection assay (RTRI) (e.g. Asante HIV-1 Rapid Recency Test or Maxim Swift Rapid Infection Assay) Identify samples that are clearly HIV negative, HIV-1 recent infection, and HIV-1 long-term infection based on the results of the RTRI Heat inactivate HIV positive specimens Set up water bath at 56 °C Incubate sample in 50ml tube at this temperature for 30 mins. Ensure sample level is completely immersed in the water bath

6 Assembling the Panel Print specimen labels indicating specimen ID, preparation date, and name of laboratory where panels are prepared Label as many 2 mL v-bottom microtubes as needed Aliquot 100 µL sample into these properly labelled microtubes and cap immediately Handle ONLY ONE specimen and ONLY ONE set of tubes at a time

Recommended Composition of QC and Training Panel 7 Recommended Composition of QC and Training Panel Verify the status of every specimen There should be 3 samples for the QC panel, and no less than 10 samples for the training panel There should be no ambiguity with the results for samples used for QC Assign specimen IDs as shown; IDs do not have to be assigned in the same order Specimen IDs HIV Status Negative Control HIV negative Recent Control HIV-1 recent Long-term Control HIV-1 long-term TP-1 TP-2 TP-3 TP-4 TP-5 TP-6 TP-7 TP-8 TP-9 TP-10

Verification of the QC and Training Panel 8 Verification of the QC and Training Panel Randomly select at least 3 sets of QCs and Training panels from the freezer and re-test using the HIV-1 RTRI 3 laboratorians should perform testing independently on all samples (if 5 sets were selected then 5 laboratorians should test) Record the visual results from all operators in the Reference Results Form

Reference Results Generation 9 Reference Results Generation LT Recent Negative Recent The final interpretation, which is the best of 3 results, will serve as the reference value for future comparisons

Panel Storage and Stability 10 Panel Storage and Stability Storage: Short-term storage (<7 days): store at 4°C. Long-term storage: store at -20°C. Stability: A maximum of 5 freeze-and-thaw cycles are acceptable. Label thawed specimens with the date removed and name/initials of the laboratory tech

Usage of the panels

Usage of the Training and QC Panel 12 Usage of the Training and QC Panel Use the Training panel during RTRI training to ascertain the competency and readiness of the trainees to provide this testing service Use the QC panel to verify performance of RTRI kits Once a month per site When storage temperature exceeds the manufacturer’s recommended storage conditions On each new lot of HIV-1 RTRI kits (NRL only) On each new shipment of HIV-1 RTRI kits (NRL only)

Record QC Results in the QC Logbook Form-Asante 13 Record QC Results in the QC Logbook Form-Asante

Record QC Results in the QC Logbook Form-Maxim Swift 14 Record QC Results in the QC Logbook Form-Maxim Swift

Review How should the specimens from the blood bank be re-tested? 15 Review How should the specimens from the blood bank be re-tested? What is the composition of the QC and training panel? How many laboratorians are needed to verify the panels? At what temperature should the panels be stored at? What is the stability of the panels? When should the panels be used?

Thank You