Prospective Evaluation of Diagnostic Accuracy of Dried Blood Spots from Finger Prick Samples for Determination of HIV-1 Load with the NucliSENS Easy-Q HIV-1 Version 2.0 Assay in Malawi

Fajardo, Emmanuel; Metcalf, Carol; Chaillet, Pascale; Aleixo, Lucia; Pannus, Pieter; Panunzi, Isabella; Triviño, Laura; Ellman, Tom; Likaka, Andrew; Mwenda, Reuben

doi:10.1128/jcm.03519-13

Cited by 27 publications

(22 citation statements)

References 26 publications

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“…However, for those DBS with a VL of over 50,000 copies/ml, they could be still a suitable specimen type. The reasons for the reduction in genotyping efficiency, in spite of the relatively insignificant impact on VL testing (36), are unclear. Possible explanations include the reduced concentration of virus particles in capillary blood due to the dilution from tissue fluid present only in fingerprick blood, presence of interfering substances or nucleases, or lower absolute volume of blood spotted onto each spot from finger-prick blood compared to the precise spotting by pipetting anticoagulated venous blood.…”

Section: Discussionmentioning

confidence: 99%

Field Study of Dried Blood Spot Specimens for HIV-1 Drug Resistance Genotyping

Parry

Parkin

Diallo³

et al. 2014

J Clin Microbiol

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h Dried blood spots (DBS) are an alternative specimen type for HIV drug resistance genotyping in resource-limited settings. Data relating to the impact of DBS storage and shipment conditions on genotyping efficiency under field conditions are limited. We compared the genotyping efficiencies and resistance profiles of DBS stored and shipped at different temperatures to those of plasma specimens collected in parallel from patients receiving antiretroviral therapy in Uganda. Plasma and four DBS cards from anti-coagulated venous blood and a fifth card from finger-prick blood were prepared from 103 HIV patients with a median viral load (VL) of 57,062 copies/ml (range, 1,081 to 2,964,191). DBS were stored at ambient temperature for 2 or 4 weeks or frozen at ؊80°C and shipped from Uganda to the United States at ambient temperature or frozen on dry ice for genotyping using a broadly sensitive in-house method. Plasma (97.1%) and DBS (98.1%) stored and shipped frozen had similar genotyping efficiencies. DBS stored frozen (97.1%) or at ambient temperature for 2 weeks (93.2%) and shipped at ambient temperature also had similar genotyping efficiencies. Genotyping efficiency was reduced for DBS stored at ambient temperature for 4 weeks (89.3%, P ‫؍‬ 0.03) or prepared from finger-prick blood and stored at ambient temperature for 2 weeks (77.7%, P < 0.001) compared to DBS prepared from venous blood and handled similarly. Resistance profiles were similar between plasma and DBS specimens. This report delineates the optimal DBS collection, storage, and shipping conditions and opens a new avenue for cost-saving ambient-temperature DBS specimen shipments for HIV drug resistance (HIVDR) surveillances in resource-limited settings.

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Section: Discussionmentioning

confidence: 99%

Field Study of Dried Blood Spot Specimens for HIV-1 Drug Resistance Genotyping

Parry

Parkin

Diallo³

et al. 2014

J Clin Microbiol

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“…Previous CD4 test evaluations have shown mixed results in terms of concordance between venous and capillary specimens 30, 31 . Numerous studies have suggested the opportunity for venous DBS for VL monitoring 27, 29, 32-44 but ours was one of the first tests of fingerstick DBS under true field conditions. We relied on existing clinic personnel for specimen collection, card preparation, and transport to the central laboratory, reflecting a more “real-world” scenario of VL monitoring using DBS via fingerstick.…”

Section: Introductionmentioning

confidence: 96%

“…DBS from fingersticks should decrease associated costs compared to venous DBS, task-shifting to lower-level providers and reducing consumable-associated expenses. Fingerstick DBS may also expand monitoring to health centers without phlebotomy capabilities 29 . Previous CD4 test evaluations have shown mixed results in terms of concordance between venous and capillary specimens 30, 31 .…”

Section: Introductionmentioning

confidence: 99%

Measures of viral load using Abbott RealTime HIV-1 Assay on venous and fingerstick dried blood spots from provider-collected specimens in Malawian District Hospitals

Rutstein

Kamwendo²,

Lebah

et al. 2014

Journal of Clinical Virology

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Background Viral suppression is a key indicator of antiretroviral therapy (ART) response among HIV-infected patients. Dried blood spots (DBS) are an appealing alternative to conventional plasma-based virologic testing, improving access to monitoring in resource-limited settings. However, validity of DBS obtained from fingerstick in field settings remains unknown. Objectives Investigate feasibility and accuracy of DBS vs plasma collected by healthcare workers in real-world settings of remote hospitals in Malawi. Compare venous DBS to fingerstick DBS for identifying treatment failure. Study design We recruited patients from ART clinics at two district hospitals in Malawi, collecting plasma, venous DBS (vDBS), and fingerstick DBS (fsDBS) cards for the first 149 patients, and vDBS and fsDBS only for the subsequent 398 patients. Specimens were tested using Abbott RealTime HIV-1 Assay (lower detection limit 40 copies/ml (plasma) and 550 copies/ml (DBS)). Results 21/149 (14.1%) had detectable viremia (>1.6 log copies/ml), 13 of which were detectable for plasma, vDBS, and fsDBS. Linear regression demonstrated high correlation for plasma vs. DBS (vDBS: β=1.19, R2 0.93 (p<0.0001); fsDBS β=1.20, R2 0.90 (p<0.0001)) and vDBS vs. fsDBS (β=0.88, R2 0.73, (p<0.0001)). Mean difference between plasma and vDBS was 0.51 log copies/ml [SD: 0.33] and plasma and fsDBS 0.46 log copies/ml [SD: 0.30]. At 5000 copies/ml, sensitivity was 100%, and specificity was 98.6% and 97.8% for vDBS and fsDBS, respectively, compared to plasma. Conclusions DBS from venipuncture and fingerstick perform well at the failure threshold of 5000 copies/ml. Fingerstick specimen source may improve access to virologic treatment monitoring in resource-limited settings given task-shifting in high-volume, low-resource facilities.

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“…One of the current limitations of measuring HIV-1 load with DBS is interference of HIV-1 DNA coamplified during HIV-1 RNA quantification by reverse transcription-PCR, notably, among samples collected from patients who were successfully treated with ART (7)(8)(9)(10)(11). Here, we determined the HIV-1 DNA level that interfered with the reliability of HIV-1 RNA quantification on DBS without using a DNase pretreatment step or a specific RNA extraction method.…”

mentioning

confidence: 99%

“…The bioMérieux NucliSENS assay is currently Conformité Européenne In Vitro Diagnostics (CE-IVD) approved for DBS specimens. Other manufacturers of HIV RNA nucleic acid tests need to pursue regulatory approval for in vitro diagnostics on DBS (6,7).…”

mentioning

confidence: 99%