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

Abstract

<jats:p> <jats:bold>Background.</jats:bold> 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.</jats:p> <jats:p> <jats:bold>Methodology.</jats:bold> 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).</jats:p> <jats:p> <jats:bold>Results.</jats:bold> In total, 107 patients were included. No statistical differences (<jats:italic>P</jats:italic>&gt;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<jats:sup>−1</jats:sup>, 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.</jats:p> <jats:p> <jats:bold>Conclusion.</jats:bold> 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.</jats:p>

Optimizing Release of Nucleic Acids of African Swine Fever Virus and Influenza A Virus from FTA Cards

“… Method 5 (M5), Nucleic acid isolation using complete lysis-M reagent (Roche Diagnostics GmbH, Mannheim, Germany): 400 µL of M-lysis reagent were added to the three punches and then incubated at 1000 rpm, 26 °C for 30 min in a thermal shaker, which was followed by centrifugation at 7000× g for 30 s. The supernatant was transferred to a new Eppendorf tube and used for both nucleic acid extraction and for the direct PCR amplification. This buffer was still successfully used for the viral RNA releasing from FTA cards [ 24 ]. Method 6 (M6-E1), Nucleic acid isolation using Chelex ® 100 Resin (Bio-Rad Laboratories, Inc., Hercules, CA, USA) and TE buffer (pH8.0, Sigma-Aldrich): 500 µL of TE buffer were added to the punches followed by the incubation at 1000 rpm, 26 °C for 30 min in a thermal shaker, followed by centrifugation at 7000× g for 30 s. The supernatant was transferred to a new Eppendorf tube and stored as eluate 1 [ 19 ].…”

“…Method 5 (M5), Nucleic acid isolation using complete lysis-M reagent (Roche Diagnostics GmbH, Mannheim, Germany): 400 µL of M-lysis reagent were added to the three punches and then incubated at 1000 rpm, 26 °C for 30 min in a thermal shaker, which was followed by centrifugation at 7000× g for 30 s. The supernatant was transferred to a new Eppendorf tube and used for both nucleic acid extraction and for the direct PCR amplification. This buffer was still successfully used for the viral RNA releasing from FTA cards [ 24 ].…”

A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies

“…Continuous improvements to the DNA extraction kit, while beneficial for the singular study, can be a problem with long-term studies when results are no longer comparable due to differences in kit effectiveness. FTA cards, on the other hand, are routinely used in governmental programmes such as the dried blood spots for newborn screening of metabolic disorders [ [44] , [45] , [46] ]. This long-term, global application of FTA cards may make them more resistant to change over time.…”