Browsing by Author "Gutema, Gadissa"

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    Effect of Specimen Handling on Quantitative HIV-1 Viral Load Measurement at Saint Paul Hospital Millennium Medical College, Addis Ababa
    (Addis Ababa University, 2019-11) Gutema, Gadissa; Tsegaye, Aster(MSc, PhD); Tola, Habteyes(MSc, PhD Fellow)
    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.