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The effect of sample handling on cross sectional HIV incidence testing results
Oliver Laeyendecker1, Amanda Latimore, Susan H Eshleman
1Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Baltimore, Maryland, United States of America. olaeyen1@jhmi.edu
Objective(S):
To determine if mishandling prior to testing would make a sample from a chronically infected subject appear recently infected when tested by cross-sectional HIV incidence assays.
Methods:
Serum samples from 31 subjects with chronic HIV infection were tested. Samples were subjected to different handling conditions, including incubation at 4 °C, 25 °C and 37 °C, for 1, 3, 7 or 15 days prior to testing. Samples were also subjected to 1,3, 7 and 15 freeze-thaw cycles prior to testing. Samples were tested using the BED capture enzyme immuno assay (BED-CEIA), Vironostika-less sensitive (V-LS), and an avidity assay using the Genetic Systems HIV-1/HIV-2 plus O EIA (avidity assay).
Results:
Compared to the sample that was not subjected to any mishandling conditions, for the BED-CEIA, V-LS and avidity assay, there was no significant change in test results for samples incubated at 4 °C or 25 °C prior to testing. No impact on test results occurred after 15 freeze-thaw cycles. A decrease in assay results was observed when samples were held for 3 days or longer at 37 °C prior to testing.
Conclusions:
Samples can be subjected up to 15 freeze-thaw cycles without affecting the results the BED-CEIA, Vironostika-LS, or avidity assays. Storing samples at 4 °C or 25 °C for up to fifteen days prior to testing had no impact on test results. However, storing samples at 37°C for three or more days did affect results obtained with these assays.
Insights
Mishandling HIV samples at room temperature or refrigerated conditions does not impact HIV incidence assay results. However, storing samples at 37°C for three days or longer can affect HIV testing outcomes.
Area of Science:
- Clinical diagnostics
- Immunology
- Public health
Background:
- Accurate HIV incidence testing is crucial for monitoring epidemics.
- Cross-sectional HIV incidence assays rely on sample integrity.
- Potential sample mishandling can affect test accuracy.
Purpose of the Study:
- To evaluate the impact of pre-testing sample mishandling on HIV incidence assay results.
- To determine if improper storage conditions can lead to misclassification of HIV infection status.
Main Methods:
- Serum samples from 31 chronically HIV-infected subjects were used.
- Samples underwent various storage conditions: 4°C, 25°C, 37°C for up to 15 days.
- Samples were also subjected to up to 15 freeze-thaw cycles.
- Testing utilized BED capture enzyme immunoassay (BED-CEIA), Vironostika-less sensitive (V-LS), and an avidity assay.
Main Results:
- Incubation at 4°C or 25°C for up to 15 days did not significantly alter BED-CEIA, V-LS, or avidity assay results.
- Up to 15 freeze-thaw cycles had no impact on the tested HIV incidence assays.
- Storage at 37°C for 3 days or longer led to decreased assay results.
Conclusions:
- HIV incidence assays (BED-CEIA, V-LS, avidity) are robust to up to 15 freeze-thaw cycles.
- Refrigerated or room temperature storage (up to 15 days) is acceptable for these HIV assays.
- Elevated temperatures (37°C) for extended periods (≥3 days) compromise the accuracy of HIV incidence testing.
