Dataset Information

Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost.

ABSTRACT: Viral load (VL) measurements are critical to the proper management of HIV in developing countries. However, access to VL assays is limited by the high cost and complexity of existing assays. While there is a need for low cost VL assays, performance must not be compromised. Thus, new assays must be validated on metrics of limit of detection (LOD), accuracy, and dynamic range. Patient plasma samples from the Joint Clinical Research Centre in Uganda were de-identified and measured using both an existing VL assay (Abbott RealTime HIV-1) and our assay, which combines low cost reagents with a simplified method of RNA isolation termed Exclusion-Based Sample Preparation (ESP).71 patient samples with VLs ranging from <40 to >3,000,000 copies/mL were used to compare the two methods. We demonstrated equivalent LOD (~50 copies/mL) and high accuracy (average difference between methods of 0.08 log, R2 = 0.97). Using expenditures from this trial, we estimate that the cost of the reagents and consumables for this assay to be approximately $5 USD. As cost is a significant barrier to implementation of VL testing, we anticipate that our assay will enhance access to this critical monitoring test in developing countries.

SUBMITTER: Berry SM

PROVIDER: S-EPMC4667969 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost.

Berry Scott M SM Pezzi Hannah M HM Williams Eram D ED Loeb Jennifer M JM Guckenberger David J DJ Lavanway Alex J AJ Puchalski Alice A AA Kityo Cissy M CM Mugyenyi Peter N PN Graziano Franklin M FM Beebe David J DJ

PloS one 20151202 12

Viral load (VL) measurements are critical to the proper management of HIV in developing countries. However, access to VL assays is limited by the high cost and complexity of existing assays. While there is a need for low cost VL assays, performance must not be compromised. Thus, new assays must be validated on metrics of limit of detection (LOD), accuracy, and dynamic range. Patient plasma samples from the Joint Clinical Research Centre in Uganda were de-identified and measured using both an exi ...[more]

PMID: 26630135

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Project description:In this study, ultrasound (US) was evaluated for As, Cd, Pb, Mn, Sr and V extraction from seaweed samples. The following parameters of ultrasound-assisted extraction (UAE) using an US bath were: frequency (25 to 130 kHz), amplitude (30 to 100%), temperature (30 to 80 °C), sample mass (50 to 200 mg), extractant concentration (1 to 3 mol L-1 of HNO3) and treatment time (5 to 30 min). Acoustic density and power density distribution were calculated using the calorimetric method and mapping of the acoustic pressure distribution was also evaluated. The optimized UAE conditions were 200 mg of sample in 10 mL of 2 mol L-1 HNO3 and 30 min of sonication in a 25 kHz US bath (37.2 ± 4.0 W L-1) at 70% of amplitude and 70 °C. Analytes were quantified using inductively coupled plasma mass spectrometry and results were compared with values obtained using "silent" conditions (magnetic or mechanical stirring at 500 rpm, and without stirring), and a reference method based on microwave-assisted wet digestion (MAWD). The UAE method demonstrated the best extraction efficiency (higher than 95%) for all analytes, especially for As, Cd and V, with lower standard deviations (up to 5%) and lower blank values in comparison with the silent conditions. The proposed UAE method was more advantageous than the reference method, being faster, simpler, safer, more environmentally friendly, and with higher detectability (lower limits of quantification, from 0.0033 to 1.34 µg g-1). In addition, negligible blank values were obtained for UAE and no interference were observed in the determination step. Furthermore, the optimized UAE method was applied for Antarctic seaweed samples and comparison with results obtained by MAWD was satisfactory. In this sense, UAE is demonstrated to be a suitable option for sample preparation of seaweed samples and further determination of environmentally critical elements avoiding the use of concentrated reagents as in the MAWD reference method.

Dataset Information

Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost.

Publications

Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost.

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