scholarly journals Whatman FTA® cards versus plasma specimens for the quantitation of HIV-1 RNA using two Real-Time PCR assays

2019 ◽  
Author(s):  
Abdourahamane Yacouba ◽  
Malika Congo ◽  
Gérard Komonsira Dioma ◽  
Hermann Somlaré ◽  
David Coulidiaty ◽  
...  
Keyword(s):  
Viral Load ◽  
Filter Paper ◽  
Load Testing ◽  
Altman Analysis ◽  
Limits Of Agreement ◽  
Bland Altman Analysis ◽  
Viral Loads ◽  
Fta Cards ◽  
Pcr Assays ◽  
Hiv 1

AbstractBackgroundSeveral studies have been conducted to compare the use DBS as alternative to plasma specimens, but mainly using Whatman 903® cards as filter paper. The aim of this study was to evaluate Whatman FTA® cards (FTA cards) specimens for HIV-1 viral load testing by comparing it to plasma specimens, using 2 real-Time PCR assays.MethodologyA cross-sectional study was conducted between April 2017 and September 2017, in HIV-1 patients admitted at Yalgado Ouédraogo teaching hospital. Paired FTA cards and plasma specimens were collected and analyzed using Abbott RealTime HIV-1 assay (Abbott) and COBAS® AmpliPrep/COBAS® TaqMan v2.0 (Roche), following manufacturers’ protocol.ResultsA total of 107 patients were included. No Statistical differences (p-value > 0.05) were observed between the mean viral loads obtained from FTA cards and plasma specimens with Roche and Abbott assays. Twenty-nine samples with Roche and 15 samples with Abbott assay showed discrepant results. At viral loads of ≤1000 copies/mL, the sensitivity and specificity of FTA cards were 78.6%, and 100% with Roche, and 92.3% and 95.9% with Abbott. Strong correlation was found between FTA cards and plasma specimens with both assays. With Roche, Bland-Altman analysis showed bias of −0.3 and 95% limits of agreement of −2.6 to 1.8 log10, with 97/99 cases (97.9%) within agreement limits. With Abbott, Bland-Altman analysis showed bias of −0.1 and 95% limits of agreement of −2.3 to 2.1 log10, with 96/99 cases (96.9%) within agreement limits.ConclusionOur study demonstrated the feasibility of using FTA cards filter paper for HIV-1 viral load testing. However, further studies are required for FTA cards filter paper validation in HIV-1 treatment monitoring.

scholarly journals Whatman FTA cards versus plasma specimens for the quantitation of HIV-1 RNA using two real-time PCR assays

2020 ◽  
Vol 2 (8) ◽  
Author(s):  
Abdourahamane Yacouba ◽  
Malika Congo ◽  
Gérard Komonsira Dioma ◽  
Hermann Somlare ◽  
David Coulidiaty ◽  
...  
Keyword(s):  
Viral Load ◽  
Real Time ◽  
Filter Paper ◽  
Real Time Pcr ◽  
Load Testing ◽  
Viral Loads ◽  
Viral Load Testing ◽  
Fta Cards ◽  
Pcr Assays ◽  
Hiv 1

Background. Several studies have compared the use of dried blot spot (DBS) as an alternative to plasma specimens, mainly using Whatman 903 cards as filter paper. The aim of this study was to evaluate the use of Whatman FTA card (FTA card) specimens for HIV-1 viral load testing compared to plasma specimens using two real-time PCR assays manufactured by Roche and Abbott. Methodology. A cross-sectional study was conducted between April 2017 and September 2017 on HIV-1 patients admitted to Yalgado Ouédraogo Teaching Hospital. Paired FTA cards and plasma specimens were collected and analysed using the Abbott Real-Time HIV-1 assay (Abbott) and COBAS AmpliPrep/COBAS TaqMan v2.0 (Roche). Results. In total, 107 patients were included. No statistical differences (P>0.05) were observed between the mean viral loads obtained from the FTA cards and those of the plasma specimens using the Roche and Abbott assays. In total, 29 samples with Roche and 15 samples with Abbott assay showed discrepant results. At viral loads of ≤1000 copies ml−1, the sensitivity and specificity of the FTA cards were 78.6 and 100% with Roche, and 92.3 and 95.9% with Abbott, respectively. Both the Roche and Abbott assays showed good correlation and agreement between the FTA cards and plasma values. Conclusion. Our study demonstrates the feasibility of using FTA card filter paper for HIV-1 viral load testing. However, further studies will be required for the validation of the use of FTA card filter paper in HIV-1 treatment monitoring.

scholarly journals Cryptococcal Antigenemia in Human Immunodeficiency Virus Antiretroviral Therapy–Experienced Ugandans With Virologic Failure

2019 ◽  
Vol 71 (7) ◽  
pp. 1726-1731 ◽  
Author(s):  
Edward Mpoza ◽  
Radha Rajasingham ◽  
Lillian Tugume ◽  
Joshua Rhein ◽  
Maria Sarah Nabaggala ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Viral Load ◽  
Antiretroviral Therapy ◽  
Cryptococcal Meningitis ◽  
Virologic Failure ◽  
Cost Effective ◽  
World Health ◽  
Load Testing ◽  
Viral Loads ◽  
Immunodeficiency Virus

Abstract Background Detectable serum or plasma cryptococcal antigen (CrAg) precedes symptomatic cryptococcal meningitis. The World Health Organization recommends CrAg screening for human immunodeficiency virus–positive persons with CD4 count <100 cells/μL initiating antiretroviral therapy (ART). However, an increasing proportion of patients with cryptococcosis are now ART experienced. Whether CrAg screening is cost-effective in those with virologic failure is unknown. Methods We retrospectively performed nationwide plasma CrAg testing among ART-experienced Ugandan adults with virologic failure (≥1000 copies/mL) using leftover plasma after viral load testing during September 2017–January 2018. For those who were CrAg positive, we obtained ART history, meningitis occurrence, and 6-month survival via medical records review. Results Among 1186 subjects with virologic failure, 35 (3.0%) were CrAg positive with median ART duration of 41 months (interquartile range, 10–84 months). Among 25 subjects with 6-month outcomes, 16 (64%) survived, 7 (28%) died, and 2 (8%) were lost. One survivor had suffered cryptococcal meningitis 2 years prior. Two others developed cryptococcal meningitis and survived. Five survivors were known to have received fluconazole. Thus, meningitis-free survival at 6 months was 61% (14/23). Overall, 91% (32/35) of CrAg-positive persons had viral load ≥5000 copies/mL compared with 64% (735/1151) of CrAg-negative persons (odds ratio, 6.0 [95% confidence interval, 1.8–19.8]; P = .001). CrAg prevalence was 4.2% (32/768) among those with viral loads ≥5000 copies/mL and 0.7% (3/419) among those with viral loads <5000 copies/mL. Conclusions In addition to the CD4 threshold of <100 cells/μL, reflexive CrAg screening should be considered in persons failing ART in Uganda with viral loads ≥5000 copies/mL.

scholarly journals Performance of a high-throughput next-generation sequencing method for analysis of HIV drug resistance and viral load

2020 ◽  
Vol 75 (12) ◽  
pp. 3510-3516 ◽  
Author(s):  
Jessica M Fogel ◽  
David Bonsall ◽  
Vanessa Cummings ◽  
Rory Bowden ◽  
Tanya Golubchik ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Viral Load ◽  
Next Generation Sequencing ◽  
High Throughput ◽  
Whole Genome ◽  
Hiv Viral Load ◽  
Viral Loads ◽  
Viral Load Assay ◽  
Generation Sequencing ◽  
Hiv 1

Abstract Objectives To evaluate the performance of a high-throughput research assay for HIV drug resistance testing based on whole genome next-generation sequencing (NGS) that also quantifies HIV viral load. Methods Plasma samples (n = 145) were obtained from HIV-positive MSM (HPTN 078). Samples were analysed using clinical assays (the ViroSeq HIV-1 Genotyping System and the Abbott RealTime HIV-1 Viral Load assay) and a research assay based on whole-genome NGS (veSEQ-HIV). Results HIV protease and reverse transcriptase sequences (n = 142) and integrase sequences (n = 138) were obtained using ViroSeq. Sequences from all three regions were obtained for 100 (70.4%) of the 142 samples using veSEQ-HIV; results were obtained more frequently for samples with higher viral loads (93.5% for 93 samples with >5000 copies/mL; 50.0% for 26 samples with 1000–5000 copies/mL; 0% for 23 samples with <1000 copies/mL). For samples with results from both methods, drug resistance mutations (DRMs) were detected in 33 samples using ViroSeq and 42 samples using veSEQ-HIV (detection threshold: 5.0%). Overall, 146 major DRMs were detected; 107 were detected by both methods, 37 were detected by veSEQ-HIV only (frequency range: 5.0%–30.6%) and two were detected by ViroSeq only. HIV viral loads estimated by veSEQ-HIV strongly correlated with results from the Abbott RealTime Viral Load assay (R2 = 0.85; n = 142). Conclusions The NGS-based veSEQ-HIV method provided results for most samples with higher viral loads, was accurate for detecting major DRMs, and detected mutations at lower levels compared with a method based on population sequencing. The veSEQ-HIV method also provided HIV viral load data.

scholarly journals Longitudinal comparison between plasma and seminal HIV-1 viral loads during antiretroviral treatment

2003 ◽  
Vol 36 (6) ◽  
pp. 689-694 ◽  
Author(s):  
Lauro Ferreira da Silva Pinto Neto ◽  
Nilo F.R. Vieira ◽  
Moacir Soprani ◽  
Carla B. Cunha ◽  
Valéria P. Cabral ◽  
...  
Keyword(s):  
Viral Load ◽  
Antiretroviral Treatment ◽  
Cd8 T Cell ◽  
Median Viral Load ◽  
Viral Loads ◽  
Cell Counts ◽  
Days Of Therapy ◽  
Longitudinal Comparison ◽  
The Impact ◽  
Hiv 1

This study was designed to investigate the impact of anti-retroviral therapy on both plasma and seminal HIV-1 viral loads and the correlation between viral loads in these compartments after treatment. Viral load, CD4+ and CD8+ T-cell counts were evaluated in paired plasma and semen samples from 36 antiretroviral therapy-naïve patients at baseline and on days 45, 90, and 180 of treatment. Slopes for blood and seminal viral loads in all treated patients were similar (p = 0.21). Median HIV-1 RNA titers in plasma and semen at baseline were 4.95 log10 and 4.48 log10 copies/ml, respectively. After 180 days of therapy, the median viral load declined to 3.15 log10 copies/ml (plasma) and 3.2 log10 copies/ml (semen). At this timepoint 22 patients presented HIV-1 viral load below 400 copies/ml in either plasma or semen, but only 9 had viral loads below 400 copies/ml in both compartments.

scholarly journals Progress in Quantitative Viral Load Testing: Variability and Impact of the WHO Quantitative International Standards

2016 ◽  
Vol 55 (2) ◽  
pp. 423-430 ◽  
Author(s):  
R. T. Hayden ◽  
Y. Sun ◽  
L. Tang ◽  
G. W. Procop ◽  
D. R. Hillyard ◽  
...  
Keyword(s):  
Viral Load ◽  
Human Herpesvirus ◽  
Bk Virus ◽  
International Standards ◽  
Load Testing ◽  
Viral Loads ◽  
Barr Virus ◽  
Viral Load Testing ◽  
Epstein Barr ◽  
The Impact

ABSTRACTIt has been hoped that the recent availability of WHO quantitative standards would improve interlaboratory agreement for viral load testing; however, insufficient data are available to evaluate whether this has been the case. Results from 554 laboratories participating in proficiency testing surveys for quantitative PCR assays of cytomegalovirus (CMV), Epstein-Barr virus (EBV), BK virus (BKV), adenovirus (ADV), and human herpesvirus 6 (HHV6) were evaluated to determine overall result variability and then were stratified by assay manufacturer. The impact of calibration to international units/ml (CMV and EBV) on variability was also determined. Viral loads showed a high degree of interlaboratory variability for all tested viruses, with interquartile ranges as high as 1.46 log10copies/ml and the overall range for a given sample up to 5.66 log10copies/ml. Some improvement in result variability was seen when international units were adopted. This was particularly the case for EBV viral load results. Variability in viral load results remains a challenge across all viruses tested here; introduction of international quantitative standards may help reduce variability and does so more or less markedly for certain viruses.

Photopheresis in HIV-1 Infected Patients (pt) using Benzoporphyrin Derivative (BPD-MA) Induces Apoptosis in CD4 Cells, Increases Intracellular Expression of Chemokines and Decreases HIV Infectivity and Viral Load.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3836-3836
Author(s):  
Zale P. Bernstein ◽  
Thomas Dougherty ◽  
Stanley A. Schwartz ◽  
Sandra Gollnick ◽  
Carleton Stewart ◽  
...  
Keyword(s):  
Viral Load ◽  
Immune System ◽  
Ex Vivo ◽  
Light Exposure ◽  
Area Under The Curve ◽  
Additional Patient ◽  
Viral Control ◽  
Viral Loads ◽  
Hiv 1

Abstract HIV is able to elude both cellular and humoral arms of the immune system; thereby making viral control difficult. Extra corporeal photochemotherapy (ECP) or photopheresis is an immunomodulatory therapy in which lymphocytes are reinfused into the host after exposure to a photoactive compound and ultraviolet A light. It is an effective therapeutic approach to several disorders of the immune system including acute and chronic graft-versus-host disease, scleroderma, and cutaneous T-cell lymphoma. This process may offer a novel approach to viral control with minimal or no toxicity. We initiated an ex vivo and subsequently a clinical pilot trial utilizing Benzoporphyrin Derivative as the photosensitive compound. Ex vivo dosing studies identified the minimum energy levels of light exposure and concentrations of BPD that eradicated both cell-free and cell-associated HIV-1 infectivity without destroying the virus particles or infected leukocytes. Leukocytes so treated remained viable. They did demonstrate altered cytokine and chemokine expression with apoptosis induced in a minority of CD4 but not CD8 positive cells. A pilot in vivo, 24 week clinical trial in seven HIV-1 infected patients (all were required, upon entry, to have viral loads of > 10,000) using the photopheresis parameters established above demonstrated that the treatment was well tolerated and beneficial. Three individuals who had rapidly rising viral loads prior to initiating therapy stabilized once treatment began. Two of which had a (sustained) greater than.5 log decrement and 5 had stable plasma viral loads (less than a.5 log increment or decrement) with varied effects on absolute CD4 and CD8 positive lymphocytes counts. One subject achieved a greater than 1 log decrement in HIV-1 plasma viral load also developed undetectable in vivo cell-free and cell-associated HIV-1 infectivity while demonstrating an increased in vitro lymphocyte mitogen stimulation index. Subsequently, under amended protocol 3 additional 12 month courses were administered to one additional patient and two of the previous enrollees. The area under the curve for viral load (viral load x # of weeks) for these ten courses of therapy showed a significant decrease from pre to post therapy (p 0.007). There were no significant changes in CD4 or CD8 numbers area under the curve (CD4 number # of weeks and CD8 number x # of weeks). None of the subjects developed an AIDS defining illness during the course of therapy nor were there any treatment associated toxicities. These studies suggest that ECP augments activity of various arms of the immune system without any significant toxicity and may be effective in controlling HIV replication. We have now instituted a Phase II study utilizing long-term photopheresis (twice monthly for 48 weeks) to further determine efficacy and mechanism of activity.

Photopheresis in HIV-1 Infected Patients (Pt) Using Benzoporphyrin Derivative (BPD-MA) Induces Apoptosis in CD4 Cells, Increases Cytolytic T-Cell Activity, Intracellular Expression of Chemokines, and Decreases HIV Infectivity and Viral Load.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1257-1257
Author(s):  
Zale P. Bernstein ◽  
Thomas Dougherty ◽  
Stanley Schwartz ◽  
Sandra Gollnick ◽  
Carleton Stewart ◽  
...  
Keyword(s):  
Viral Load ◽  
Immune System ◽  
T Cell ◽  
Ex Vivo ◽  
Cell Activity ◽  
Area Under The Curve ◽  
Viral Control ◽  
Viral Loads ◽  
Hiv 1

Abstract HIV is able to elude both cellular and humoral arms of the immune system; thereby making viral control difficult. Extra corporeal photochemotherapy (ECP) or photopheresis is an immunomodulatory therapy in which lymphocytes are reinfused into the host after exposure to a photoactive compound and ultraviolet A light. It is an effective therapeutic approach to several disorders of the immune system including acute and chronic graft-versus-host disease, scleroderma, and cutaneous T-cell lymphoma. This process may offer a novel approach to viral control with minimal or no toxicity. We initiated an ex vivo and subsequently a clinical pilot trial utilizing Benzoporphyrin Derivative as the photosensitive compound. Ex vivo dosing studies identified the minimum energy levels of light exposure and concentrations of BPD that eradicated both cell-free and cell-associated HIV-1 infectivity without destroying the virus particles or infected leukocytes. Leukocytes so treated remained viable. They did demonstrate altered cytokine and chemokine expression with apoptosis induced in a minority of CD4 but not CD8 positive cells. Furthermore, there was a statistically significant increase in cytolytic T-cell activity expressed as percentage of granzyme-B release. A pilot in vivo, 24 week clinical trial in seven HIV-1 infected patients (all were required, upon entry, to have viral loads of > 10,000) using the photopheresis parameters established above demonstrated that the treatment was well tolerated and beneficial. Three individuals who had rapidly rising viral loads prior to initiating therapy stabilized once treatment began. Two of which had a (sustained) greater than .5 log decrement and 5 had stable plasma viral loads (less than a .5 log increment or decrement) with varied effects on absolute CD4 and CD8 positive lymphocytes counts. One subject achieved a greater than 1 log decrement in HIV-1 plasma viral load also developed undetectable in vivo cell-free and cell-associated HIV-1 infectivity while demonstrating an increased in vitro lymphocyte mitogen stimulation index. Subsequently, under amended and successor protocol 5 additional 12 month courses were administered to three additional patients and two of the previous enrollees. The area under the curve for viral load (viral load x # of weeks) for these twelve courses of therapy showed a significant decrease from pre to post therapy (p 0.007). There were no significant changes in CD4 or CD8 numbers area under the curve (CD4 number # of weeks and CD8 number x # of weeks). None of the subjects developed an AIDS defining illness during the course of therapy nor were there any treatment associated toxicities. These studies suggest that ECP augments activity of various arms of the immune system without any significant toxicity and may be effective in controlling HIV replication. We now plan a randomized Phase II study utilizing long-term photopheresis (twice monthly for 48 weeks) in addition to anti-retroviral therapy versus anti-retroviral therapy alone to further determine efficacy and mechanism of activity.

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