{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T04:01:29Z","timestamp":1791000089469,"version":"4.1.0"},"reference-count":74,"publisher":"National Academy of Sciences","issue":"7","license":[{"start":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T00:00:00Z","timestamp":1739145600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"HHS | NIH","doi-asserted-by":"publisher","award":["U54EB027049"],"award-info":[{"award-number":["U54EB027049"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010558","name":"Office of the Vice Provost for Research, Tufts University","doi-asserted-by":"publisher","award":["n\/a"],"award-info":[{"award-number":["n\/a"]}],"id":[{"id":"10.13039\/100010558","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,2,18]]},"abstract":"<jats:p>Quantifying viral load, a key indicator required to achieve control and elimination of the HIV epidemic, requires cell-free plasma or serum to ensure measurements are not biased by proviral DNA contained in infected CD4 T lymphocytes. Plasma separation cards (PSC) collect and preserve a dried specimen, which makes them practical solutions for decentralized sample collection and transport in limited-resource settings. However, physiological variations in hematocrit levels can introduce significant variability in the quality of plasma generated by commercial PSCs and can lead to inaccurate test results and clinical decisions. In addition to hematocrit-dependent sampling, the Roche PSC, a standard for dried plasma collection, is known to induce considerable hemolysis, which further impacts specimen quality, concordance with liquid plasma, and the overall benefit of microsampling. We address these gaps with a patterned dried plasma spot (pDPS) card, which generates plasma with improved hematocrit independence and minimal hemolysis. This study directly compares pDPS cards to the Roche PSC to measure HIV viral load. Analysis of viral load from 75 donors revealed strong agreement in sensitivity, specificity, overall accuracy, and viral load band placement between devices, with quantitative metrics suggesting improved performance for pDPS cards. In reflexive genotyping, remnant dried blood from pDPS cards exhibited greater success than Roche PSC in amplification and sequencing (71% vs. 62%) and detecting drug resistance mutations (63% vs. 42%). Based on this performance, pDPS cards can be versatile across multiple analytical platforms, integrate seamlessly into existing clinical laboratory workflows, and aid clinicians in making accurate treatment decisions.<\/jats:p>","DOI":"10.1073\/pnas.2419160122","type":"journal-article","created":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T15:04:36Z","timestamp":1739199876000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":7,"title":["Clinical evaluation of patterned dried plasma spot cards to support quantification of HIV viral load and reflexive genotyping"],"prefix":"10.1073","volume":"122","author":[{"given":"Giorgio Gianini","family":"Morbioli","sequence":"first","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith R.","family":"Baillargeon","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9877-6805","authenticated-orcid":false,"given":"Monalisa N.","family":"Kalimashe","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vibha","family":"Kana","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7982-5539","authenticated-orcid":false,"given":"Hloniphile","family":"Zwane","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7897-4671","authenticated-orcid":false,"given":"Cheri","family":"van der Walt","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison J.","family":"Tierney","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea C.","family":"Mora","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0420-046X","authenticated-orcid":false,"given":"Mark","family":"Goosen","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rivashni","family":"Jagaroo","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica C.","family":"Brooks","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2029-9236","authenticated-orcid":false,"given":"Ewald\u00e9","family":"Cutler","sequence":"additional","affiliation":[{"name":"Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases","place":["Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gillian","family":"Hunt","sequence":"additional","affiliation":[{"name":"Bio Analytical Research Corporation South Africa","place":["Richmond, Johannesburg, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael R.","family":"Jordan","sequence":"additional","affiliation":[{"name":"Department of Medicine, Tufts University School of Medicine","place":["Boston"]},{"name":"Department of Public Health and Community Medicine, Tufts University School of Medicine","place":["Boston"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3968-6391","authenticated-orcid":false,"given":"Alice","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Public Health and Community Medicine, Tufts University School of Medicine","place":["Boston"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3410-5014","authenticated-orcid":false,"given":"Charles R.","family":"Mace","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Laboratory for Living Devices, Tufts University","place":["Medford"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,2,10]]},"reference":[{"key":"e_1_3_4_1_2","unstructured":"UNAIDS Understanding fast-track: Accelerating action to end the AIDS epidemic by 2030. https:\/\/www.unaids.org\/en\/resources\/documents\/2015\/201506_JC2743_Understanding_FastTrack. Accessed 1 August 2024."},{"key":"e_1_3_4_2_2","unstructured":"US Centers for Disease Control and Prevention HIV treatment as prevention. https:\/\/www.cdc.gov\/hiv\/risk\/art\/index.html. Accessed 1 August 2024."},{"key":"e_1_3_4_3_2","unstructured":"UNAIDS The path that ends AIDS: 2023 UNAIDS Global AIDS update. https:\/\/thepath.unaids.org\/wp-content\/themes\/unaids2023\/assets\/files\/2023_report.pdf. Accessed 1 August 2024."},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1136\/archdischild-2013-305548"},{"key":"e_1_3_4_5_2","unstructured":"UNAIDS Global HIV statistics. https:\/\/www.unaids.org\/sites\/default\/files\/media_asset\/UNAIDS_FactSheet_en.pdf. Accessed 1 August 2024."},{"key":"e_1_3_4_6_2","unstructured":"Clinical Info HIV.gov HIV\/AIDS glossary: Viral load test. https:\/\/clinicalinfo.hiv.gov\/en\/glossary\/viral-load-test. Accessed 1 August 2024."},{"key":"e_1_3_4_7_2","unstructured":"World Health Organization HIV molecular diagnostics toolkit to Improve access to viral load testing and infant diagnosis. https:\/\/iris.who.int\/bitstream\/handle\/10665\/325961\/9789241516211-eng.pdf?sequence=1. Accessed 1 August 2024."},{"key":"e_1_3_4_8_2","unstructured":"World Health Organization What\u2019s new in treatment monitoring: viral load and CD4 testing. https:\/\/apps.who.int\/iris\/bitstream\/handle\/10665\/255891\/WHO-HIV-2017.22-eng.pdf. Accessed 1 August 2024."},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S2352-3018(23)00228-X"},{"key":"e_1_3_4_10_2","unstructured":"World Health Organization HIV drug resistance\u2013A brief report. (2024). https:\/\/www.who.int\/publications\/i\/item\/9789240086319. Accessed 1 August 2024."},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.15585\/mmwr.mm7021a3"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/btm2.10476"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01336-18"},{"key":"e_1_3_4_14_2","first-page":"1","article-title":"Performance of a novel low-cost, instrument-free plasma separation device for HIV viral load quantification and determination of treatment failure in people living with HIV in Malaysia: A diagnostic accuracy study","volume":"57","author":"Pham M. D.","year":"2018","unstructured":"M. D. Pham , Performance of a novel low-cost, instrument-free plasma separation device for HIV viral load quantification and determination of treatment failure in people living with HIV in Malaysia: A diagnostic accuracy study. J. Clin. Microbiol. 57, 1\u20139 (2018).","journal-title":"J. Clin. Microbiol."},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1111\/cts.13411"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2020.461731"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciz338"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1542\/peds.32.3.338"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2014.06.142"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dew114"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.4103\/0972-9062.373616"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11090830"},{"key":"e_1_3_4_23_2","unstructured":"World Health Organization Consolidated guidelines on the use of antiretroviral drugs for treating and preventing HIV infection: recommendations for a public health approach 2nd ed. https:\/\/www.who.int\/publications\/i\/item\/9789241549684. Accessed 1 August 2024."},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsmeasuresciau.2c00034"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.9b00204"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/hvaa082"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics10121032"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/hvab170"},{"key":"e_1_3_4_29_2","first-page":"1350","article-title":"Microfluidic device for patient-centric multiplexed assays with readout in centralized laboratories","volume":"95","author":"Hauser J.","year":"2023","unstructured":"J. Hauser , Microfluidic device for patient-centric multiplexed assays with readout in centralized laboratories. Anal. Chem. 95, 1350\u20131358 (2023).","journal-title":"Anal. Chem."},{"key":"e_1_3_4_30_2","unstructured":"Roche Diagnostics cobas Plasma Separation Card. https:\/\/diagnostics.roche.com\/global\/en\/products\/params\/cobas-plasma-separation-card.html. Accessed 1 August 2024."},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01336-18"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcv.2021.104839"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgph.0002099"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.00546-23"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.4155\/bio-2023-0057"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics11030473"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsmeasuresciau.1c00031"},{"key":"e_1_3_4_38_2","author":"World Health Organization","year":"2023","unstructured":"World Health Organization, The Role of HIV Viral Suppression in Improving Individual Health and Reducing Transmission: Policy Brief (2023).","journal-title":"The Role of HIV Viral Suppression in Improving Individual Health and Reducing Transmission: Policy Brief"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1063\/1.4790819"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1039\/C4LC00920G"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7LC01300K"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1142\/S2339547817200011"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac300689s"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1039\/C6LC00895J"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.3c04439"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.0c04127"},{"key":"e_1_3_4_47_2","unstructured":"Spot-On Sciences HemaSpot SE. https:\/\/www.spotonsciences.com\/hemaspot-se. Accessed 1 August 2024."},{"key":"e_1_3_4_48_2","author":"Geneva: World Health Organization","year":"2020","unstructured":"Geneva: World Health Organization, WHO manual for HIV drug resistance testing using dried blood spot specimens, third edition (2020).","journal-title":"WHO manual for HIV drug resistance testing using dried blood spot specimens, third edition"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2018.10.010"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.4155\/bio.11.220"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/6.3.223"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1136\/bmj.330.7497.949"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-018-0931-z"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1093\/jalm\/jfab156"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/aqab217"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1080\/10408363.2021.1943302"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7AY00505A"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/25.3.432"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1097\/QAI.0000000000002321"},{"key":"e_1_3_4_60_2","unstructured":"World Health Organization Target product profile for HIV drug resistance tests in low- and middle-income contries: Africa. https:\/\/iris.who.int\/bitstream\/handle\/10665\/372838\/9789240076662-eng.pdf?sequence=1. Accessed 1 August 2024."},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1086\/503914"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0004724"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkz417"},{"key":"e_1_3_4_64_2","first-page":"185","article-title":"Evaluation of factors influencing precision in the analysis of samples taken from blood spots on filter paper","volume":"17","author":"O\u2019Broin S. D.","year":"1995","unstructured":"S. D. O\u2019Broin, B. P. Kelleher, E. Gunter, Evaluation of factors influencing precision in the analysis of samples taken from blood spots on filter paper. Clin. Lab. Haematol. 17, 185\u2013188 (1995).","journal-title":"Clin. Lab. Haematol."},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.4155\/bio.10.103"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.9b03179"},{"key":"e_1_3_4_67_2","unstructured":"M. Pekurovsky \u201cDiagnostic device.\u201d US Patent 20230138304 (2023)."},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10404-014-1340-z"},{"key":"e_1_3_4_69_2","unstructured":"R Core Team R: A Language and Environment for Statistical Computing (version 4.3.3) (R Foundation for Statistical Computing Vienna Austria 2022). https:\/\/www.r-project.org\/."},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2015.246280"},{"key":"e_1_3_4_71_2","author":"Geneva: World Health Organization","year":"2021","unstructured":"Geneva: World Health Organization, Consolidated Guidelines on HIV Prevention, Testing, Treatment, Service Delivery and Monitoring: Recommendations for a Public Health Approach (2021).","journal-title":"Consolidated Guidelines on HIV Prevention, Testing, Treatment, Service Delivery and Monitoring: Recommendations for a Public Health Approach"},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2004.031823"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msy096"},{"key":"e_1_3_4_74_2","doi-asserted-by":"crossref","unstructured":"G. G. Morbioli Data from \u201cClinical evaluation of patterned dried plasma spot cards to support quantification of HIV viral load and reflexive genotyping.\u201d Zenodo. https:\/\/doi.org\/10.5281\/zenodo.13610433. Deposited 30 August 2024.","DOI":"10.26434\/chemrxiv-2024-5bqm7"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2419160122","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T15:04:52Z","timestamp":1739199892000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2419160122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,10]]},"references-count":74,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2025,2,18]]}},"alternative-id":["10.1073\/pnas.2419160122"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2419160122","relation":{"has-preprint":[{"id-type":"doi","id":"10.26434\/chemrxiv-2024-5bqm7","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,10]]},"assertion":[{"value":"2024-09-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-01-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-02-10","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2419160122"}}