Effect of Specimen Handling on Quantitative HIV-1 Viral Load Measurement at Saint Paul Hospital Millennium Medical College, Addis Ababa
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Date
2019-11
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Addis Ababa University
Abstract
Background: Proper specimen handling prior to quantification of plasma HIV-1 RNA
viral load is important, since some reports suggest that variations in specimen handling
may affect detection and quantification of plasma RNA. However, there is limited
evidence on the effect of time of plasma separation, storage, freeze-thawing and dilution
on the HIV-1 RNA viral load.
Objective: To determine the effect of sample management on HIV-1 RNA viral load
measurement in St. Paul Hospital Millennium Medical College from April to July 2019.
Methods: Experimental study design was conducted in St. Paul Hospital Millennium
Medical College from April to July 2019 GC in people living with HIV. Whole blood
sample was collected into two EDTA test tubes from 88 eligible participants. The viral
load test was done by Abbott m2000sp/rt analyzer. Data was entered into Microsoft excel
and analyzed by SPSS version 20. Repeated measure analysis of variance was used to
compare HIV RNA viral load mean difference between different time of plasma
separation, storage, freeze-thawing and dilution. Post-hock analysis was employed to
locate the place of significant difference. Level of significance was set at 5%.
Results: There was significant HIV-1 RNA viral load log mean difference between plasma
separation time at 6 hours (hrs) and 24hrs (p<0.001). Similarly, there was significant HIV-
1 RNA viral load log mean difference between plasma tested within 6hrs, stored at 2-80c
for 6 and 15 days (p = 0.006 and <0.001). HIV-1 RNA viral load log mean difference was
not observed when plasma was stored at 2-80c for 6 days (p= 0.999) and at - 200c for either
30 (p = 0.899) or 60 days (p = 0.999). There was significant log mean difference between
plasma that exposed to 4th cycle of freeze-thawing in -200c when compared with plasma
tested within 6hrs (p = 0.013). For the three dilution proportions (1:2, 1:3 and 1:5) there
was no significant difference on mean RNA copies when compared to each other and
tested within 6hrs (p = 0.999).
Conclusion and recommendation: Only plasma separated at 24hrs, stored at 2-8oc for
15days and freeze-thaw 4th cycle had statistically significant effect on HIV RNA viral load
variation. Though the differences were not clinically significant at a cut-off viral load level
of 0.5 log10, therefore focusing on these factors could improve the result quality and
ultimately improve patient care.
Description
This is to certify that the thesis prepared by Gadissa Gutema, entitled:
The Effect of Sample Management on Quantitative HIV-1 Viral Load Measurement at St
Paul Hospital Millennium Medical College, Addis Ababa and submitted in partial
fulfillment of the requirements for Master of Science degree in Clinical Laboratory
Sciences (Diagnostic and Public Health Microbiology) complies with the regulations of the
University and meets the accepted standards with respect to originality and quality
Keywords
Human immunodeficiency virus, HIV-1 RNA viral load, quantitative RT PCR, Abbott m2000sp/rt analyzer, Ethiopia