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 RNAviral load is important, since some reports suggest that variations in specimen handlingmay affect detection and quantification of plasma RNA. However, there is limitedevidence on the effect of time of plasma separation, storage, freeze-thawing and dilutionon the HIV-1 RNA viral load.
Objective: To determine the effect of sample management on HIV-1 RNA viral loadmeasurement in St. Paul Hospital Millennium Medical College from April to July 2019.
Methods: Experimental study design was conducted in St. Paul Hospital MillenniumMedical College from April to July 2019 GC in people living with HIV. Whole bloodsample was collected into two EDTA test tubes from 88 eligible participants. The viralload test was done by Abbott m2000sp/rt analyzer. Data was entered into Microsoft exceland analyzed by SPSS version 20. Repeated measure analysis of variance was used tocompare HIV RNA viral load mean difference between different time of plasmaseparation, storage, freeze-thawing and dilution. Post-hock analysis was employed tolocate 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 plasmaseparation 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 2-80cfor 6 and 15 days (p = 0.006 and <0.001). HIV-1 RNA viral load log mean difference wasnot observed when plasma was stored at 2-80c for 6 days (p= 0.999) and at - 200c for either30 (p = 0.899) or 60 days (p = 0.999). There was significant log mean difference betweenplasma that exposed to 4th cycle of freeze-thawing in -200c when compared with plasmatested within 6hrs (p = 0.013). For the three dilution proportions (1:2, 1:3 and 1:5) therewas no significant difference on mean RNA copies when compared to each other andtested within 6hrs (p = 0.999).
Conclusion and recommendation: Only plasma separated at 24hrs, stored at 2-8oc for15days and freeze-thaw 4th cycle had statistically significant effect on HIV RNA viral loadvariation. Though the differences were not clinically significant at a cut-off viral load levelof 0.5 log10, therefore focusing on these factors could improve the result quality andultimately improve patient care.
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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 StPaul Hospital Millennium Medical College, Addis Ababa and submitted in partialfulfillment of the requirements for Master of Science degree in Clinical LaboratorySciences (Diagnostic and Public Health Microbiology) complies with the regulations of theUniversity 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