{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T11:23:29Z","timestamp":1772623409275,"version":"3.50.1"},"reference-count":55,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2024,11,25]],"date-time":"2024-11-25T00:00:00Z","timestamp":1732492800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosglobalpublichealth.org"],"crossmark-restriction":false},"short-container-title":["PLOS Glob Public Health"],"abstract":"<jats:p>Lateral flow rapid diagnostic tests (RDTs, RTs) are cost-effective with low infrastructure requirements for limited-resource settings, and in any setting can represent a bridge between early disease monitoring at outbreak onset and fully-scaled molecular testing for human or animal diseases. However, the potential of RTs to handle higher throughput testing is hampered by the need for manual processing. Here we review dengue virus and African swine fever virus rapid tests, and present a novel protocol that employs an open-source fluid handler to automate the execution of up to 42 RTs per run. A publicly accessible website, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/rtWIZARD.lji.org\" xlink:type=\"simple\">rtWIZARD.lji.org<\/jats:ext-link>, provides printouts for correctly spacing cassettes, worksheets for sample organization, and test-specific fluid handler protocols to accurately deliver samples from a 48-tube rack to each cassette\u2019s sample and running buffer wells. An optional QR-coded sheet allows for de-identified sample-to-result traceability by producing a unique printable label for each cassette, enabling results to be entered via a scanner. This work describes a highly cost-effective model for increasing outbreak diagnostic efficiency and of increasing RT throughput for other applications including workplace testing, food safety, environmental testing, and defense applications.<\/jats:p>","DOI":"10.1371\/journal.pgph.0002625","type":"journal-article","created":{"date-parts":[[2024,11,25]],"date-time":"2024-11-25T18:41:39Z","timestamp":1732560099000},"page":"e0002625","update-policy":"https:\/\/doi.org\/10.1371\/journal.pgph.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Automation for lateral flow rapid tests: Protocol for an open-source fluid handler and applications to dengue and African swine fever tests"],"prefix":"10.1371","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-2247-166X","authenticated-orcid":true,"given":"Rohan","family":"Laurent","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benjamin","family":"Hinnant","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Talbott","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5993-4575","authenticated-orcid":true,"given":"Kenneth","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2024,11,25]]},"reference":[{"issue":"1","key":"pgph.0002625.ref001","doi-asserted-by":"crossref","first-page":"3096","DOI":"10.1038\/s41467-023-38868-w","article-title":"Highly lethal genotype I and II recombinant African swine fever viruses detected in pigs","volume":"14","author":"D Zhao","year":"2023","journal-title":"Nat Commun"},{"issue":"10","key":"pgph.0002625.ref002","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1038\/s43016-021-00362-1","article-title":"African swine fever outbreaks in China led to gross domestic product and economic losses","volume":"2","author":"S You","year":"2021","journal-title":"Nat Food"},{"key":"pgph.0002625.ref003","doi-asserted-by":"crossref","first-page":"e2021070","DOI":"10.4178\/epih.e2021070","article-title":"Dengue and COVID-19 infections in the ASEAN region: a concurrent outbreak of viral diseases","volume":"43","author":"L Wiyono","year":"2021","journal-title":"Epidemiol Health"},{"key":"pgph.0002625.ref004","unstructured":"Dengue\u2014PAHO\/WHO | Pan American Health Organization [Internet]. [cited 2023 Aug 31]. Available from: https:\/\/www.paho.org\/en\/topics\/dengue."},{"key":"pgph.0002625.ref005","unstructured":"Dengue\u2013the Region of the Americas [Internet]. [cited 2023 Aug 31]. Available from: https:\/\/www.who.int\/emergencies\/disease-outbreak-news\/item\/2023-DON475."},{"key":"pgph.0002625.ref006","article-title":"Dengue is breaking records in the Americas\u2014what\u2019s behind the surge?","author":"M. Lenharo","year":"2023","journal-title":"Nature"},{"key":"pgph.0002625.ref007","unstructured":"Ongoing avian influenza outbreaks in animals pose risk to humans [Internet]. AnimalHealth. [cited 2024 Jul 10]. Available from: https:\/\/www.fao.org\/animal-health\/news-events\/news\/detail\/ongoing-avian-influenza-outbreaks-in-animals-pose-risk-to-humans\/en."},{"key":"pgph.0002625.ref008","unstructured":"USDA. 2022\u20132024 Detections of Highly Pathogenic Avian Influenza [Internet]. [cited 2024 Jul 9]. Available from: https:\/\/www.aphis.usda.gov\/livestock-poultry-disease\/avian\/avian-influenza\/hpai-detections."},{"issue":"7","key":"pgph.0002625.ref009","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.3201\/eid3007.240508","article-title":"Highly Pathogenic Avian Influenza A(H5N1) Clade 2.3.4.4b Virus Infection in Domestic Dairy Cattle and Cats, United States, 2024","volume":"30","author":"ER Burrough","year":"2024","journal-title":"Emerg Infect Dis"},{"issue":"21","key":"pgph.0002625.ref010","doi-asserted-by":"crossref","first-page":"501","DOI":"10.15585\/mmwr.mm7321e1","article-title":"Outbreak of Highly Pathogenic Avian Influenza A(H5N1) Viruses in U.S. Dairy Cattle and Detection of Two Human Cases\u2014United States, 2024","volume":"73","author":"S Garg","year":"2024","journal-title":"MMWR Morb Mortal Wkly Rep"},{"key":"pgph.0002625.ref011","unstructured":"HPAI Detections in Mammals [Internet]. [cited 2024 Jul 10]. Available from: https:\/\/www.aphis.usda.gov\/livestock-poultry-disease\/avian\/avian-influenza\/hpai-detections\/mammals."},{"key":"pgph.0002625.ref012","unstructured":"COVID-19 deaths | WHO COVID-19 dashboard [Internet]. datadot. [cited 2024 Jul 10]. Available from: https:\/\/data.who.int\/dashboards\/covid19\/cases."},{"issue":"1","key":"pgph.0002625.ref013","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1038\/s41467-022-30698-6","article-title":"Transmission of SARS-CoV-2 from humans to animals and potential host adaptation","volume":"13","author":"CCS Tan","year":"2022","journal-title":"Nat Commun"},{"issue":"8","key":"pgph.0002625.ref014","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1016\/S1473-3099(16)00146-8","article-title":"The global economic burden of dengue: a systematic analysis","volume":"16","author":"DS Shepard","year":"2016","journal-title":"Lancet Infect Dis"},{"key":"pgph.0002625.ref015","doi-asserted-by":"crossref","first-page":"101122","DOI":"10.1016\/j.nmni.2023.101122","article-title":"The resurgence of Avian influenza and human infection: A brief outlook","volume":"53","author":"RA Farahat","year":"2023","journal-title":"New Microbes New Infect"},{"key":"pgph.0002625.ref016","unstructured":"Bird Flu\u2019s Surge Has Scientists Seeking Clues to Prevent the Next Pandemic. Bloomberg.com [Internet]. 2023 May 1 [cited 2023 Aug 31]; Available from: https:\/\/www.bloomberg.com\/news\/features\/2023-05-01\/bird-flu-surge-has-scientists-seeking-clues-to-prevent-the-next-pandemic."},{"key":"pgph.0002625.ref017","unstructured":"Avian influenza outbreaks reduced egg production, driving prices to record highs in 2022 [Internet]. [cited 2023 Sep 11]. Available from: http:\/\/www.ers.usda.gov\/data-products\/chart-gallery\/gallery\/chart-detail\/?chartId=105576."},{"key":"pgph.0002625.ref018","unstructured":"COVID-19\u2019s Total Cost to the U.S. Economy Will Reach $14 Trillion by End of 2023 [Internet]. USC Schaeffer. 2023 [cited 2023 Aug 31]. Available from: https:\/\/healthpolicy.usc.edu\/article\/covid-19s-total-cost-to-the-economy-in-us-will-reach-14-trillion-by-end-of-2023-new-research\/."},{"key":"pgph.0002625.ref019","doi-asserted-by":"crossref","unstructured":"Sands RA Gita Gopinath,Jeremy Farrar,Richard Hatchett,Peter. A Global Strategy to Manage the Long-Term Risks of COVID-19 [Internet]. IMF. [cited 2024 Jul 10]. Available from: https:\/\/www.imf.org\/en\/Publications\/WP\/Issues\/2022\/04\/04\/A-Global-Strategy-to-Manage-the-Long-Term-Risks-of-COVID-19-516079.","DOI":"10.5089\/9798400205996.001"},{"key":"pgph.0002625.ref020","doi-asserted-by":"crossref","first-page":"106147","DOI":"10.1016\/j.econmod.2022.106147","article-title":"Macroeconomic consequences of the COVID-19 pandemic","volume":"120","author":"T Walmsley","year":"2023","journal-title":"Econ Model"},{"issue":"3","key":"pgph.0002625.ref021","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1038\/s41579-020-00461-z","article-title":"Considerations for diagnostic COVID-19 tests","volume":"19","author":"O Vandenberg","year":"2021","journal-title":"Nat Rev Microbiol"},{"key":"pgph.0002625.ref022","doi-asserted-by":"crossref","first-page":"100092","DOI":"10.1016\/j.bea.2023.100092","article-title":"Medical diagnostic value of digital PCR (dPCR): A systematic review","volume":"6","author":"S. Nazir","year":"2023","journal-title":"Biomedical Engineering AdvancesNov 1"},{"issue":"7101","key":"pgph.0002625.ref023","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1038\/nature05064","article-title":"Microfluidic diagnostic technologies for global public health","volume":"442","author":"P Yager","year":"2006","journal-title":"Nature"},{"key":"pgph.0002625.ref024","doi-asserted-by":"crossref","first-page":"549","DOI":"10.3389\/fimmu.2019.00549","article-title":"Infectious Disease Threats in the Twenty-First Century: Strengthening the Global Response","volume":"10","author":"DE Bloom","year":"2019","journal-title":"Front Immunol"},{"key":"pgph.0002625.ref025","unstructured":"Ward D. Faster, Better, Cheaper Revisited: Program Management Lessons from NASA. In 2010 [cited 2023 Sep 5]. Available from: https:\/\/www.semanticscholar.org\/paper\/Faster%2C-Better%2C-Cheaper-Revisited%3A-Program-Lessons-Ward\/c52db9aed0274b725bfb24b13d05ad940c6dbae6."},{"issue":"6","key":"pgph.0002625.ref026","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1093\/jat\/bkz048","article-title":"Oral Fluid Drug Testing: Analytical Approaches, Issues and Interpretation of Results","volume":"43","author":"NA Desrosiers","year":"2019","journal-title":"J Anal Toxicol"},{"issue":"2","key":"pgph.0002625.ref027","doi-asserted-by":"crossref","first-page":"468","DOI":"10.5740\/jaoacint.17-0221","article-title":"Detection of Paralytic Shellfish Toxins in Mussels and Oysters Using the Qualitative Neogen Lateral-Flow Immunoassay: An Interlaboratory Study","volume":"101","author":"JJ Dorantes-Aranda","year":"2018","journal-title":"J AOAC Int"},{"issue":"12","key":"pgph.0002625.ref028","doi-asserted-by":"crossref","first-page":"2637","DOI":"10.4315\/0362-028X-68.12.2637","article-title":"Evaluation of detection methods for screening meat and poultry products for the presence of foodborne pathogens","volume":"68","author":"VM Bohaychuk","year":"2005","journal-title":"J Food Prot"},{"issue":"11","key":"pgph.0002625.ref029","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.3390\/bios12111002","article-title":"Development of Lateral Flow Test-System for the Immunoassay of Dibutyl Phthalate in Natural Waters","volume":"12","author":"AN Berlina","year":"2022","journal-title":"Biosensors (Basel)"},{"issue":"1","key":"pgph.0002625.ref030","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1021\/acs.chas.0c00078","article-title":"Surface Contamination Generated by \u201cOne-Pot\u201d Methamphetamine Production","volume":"28","author":"AL Ciesielski","year":"2020","journal-title":"J Chem Health Saf"},{"issue":"1","key":"pgph.0002625.ref031","doi-asserted-by":"crossref","first-page":"16157","DOI":"10.1038\/s41598-018-34610-5","article-title":"Uranium (VI) detection in groundwater using a gold nanoparticle\/paper-based lateral flow device","volume":"8","author":"D Quesada-Gonz\u00e1lez","year":"2018","journal-title":"Sci Rep"},{"issue":"2","key":"pgph.0002625.ref032","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1111\/1556-4029.14631","article-title":"The use of lateral flow immunoassays for the detection of fentanyl in seized drug samples and postmortem urine","volume":"66","author":"DJ Angelini","year":"2021","journal-title":"J Forensic Sci"},{"key":"pgph.0002625.ref033","doi-asserted-by":"crossref","first-page":"109610","DOI":"10.1016\/j.drugalcdep.2022.109610","article-title":"Validation of a lateral flow chromatographic immunoassay for the detection of fentanyl in drug samples","volume":"240","author":"JN Park","year":"2022","journal-title":"Drug Alcohol Depend"},{"issue":"3","key":"pgph.0002625.ref034","doi-asserted-by":"crossref","first-page":"402","DOI":"10.3390\/bios13030402","article-title":"Immunosensors for Assay of Toxic Biological Warfare Agents","volume":"13","author":"M. Pohanka","year":"2023","journal-title":"Biosensors (Basel)"},{"issue":"1","key":"pgph.0002625.ref035","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1186\/s12985-022-01904-0","article-title":"Performance of the nonstructural 1 Antigen Rapid Test for detecting all four DENV serotypes in clinical specimens from Bangkok","volume":"19","author":"K Poltep","year":"2022","journal-title":"Thailand. Virology Journal"},{"issue":"2","key":"pgph.0002625.ref036","doi-asserted-by":"crossref","first-page":"e0009065","DOI":"10.1371\/journal.pntd.0009065","article-title":"High performance dengue virus antigen-based serotyping-NS1-ELISA (plus): A simple alternative approach to identify dengue virus serotypes in acute dengue specimens","volume":"15","author":"T Prommool","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"pgph.0002625.ref037","doi-asserted-by":"crossref","first-page":"e02796","DOI":"10.1128\/spectrum.02796-22","article-title":"Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices","volume":"11","author":"NM Pollak","year":"2023","journal-title":"Microbiology Spectrum"},{"key":"pgph.0002625.ref038","article-title":"Development of Novel Dengue NS1 Multiplex Lateral Flow Immunoassay to Differentiate Serotypes in Serum of Acute Phase Patients and Infected Mosquitoes","author":"SC Lai","year":"2022","journal-title":"Frontiers in Immunology [Internet]"},{"issue":"9","key":"pgph.0002625.ref039","doi-asserted-by":"crossref","first-page":"e0239710","DOI":"10.1371\/journal.pone.0239710","article-title":"Evaluation of rapid diagnostic tests to detect dengue virus infections in Taiwan","volume":"15","author":"LT Liu","year":"2020","journal-title":"PLOS ONE"},{"issue":"2","key":"pgph.0002625.ref040","doi-asserted-by":"crossref","first-page":"115116","DOI":"10.1016\/j.diagmicrobio.2020.115116","article-title":"Diagnostic accuracy of 5 different brands of dengue virus non-structural protein 1 (NS1) antigen rapid diagnostic tests (RDT) in Indonesia","volume":"98","author":"MS Santoso","year":"2020","journal-title":"Diagnostic Microbiology and Infectious Disease"},{"issue":"3","key":"pgph.0002625.ref041","doi-asserted-by":"crossref","first-page":"436","DOI":"10.3201\/eid1503.080923","article-title":"Evaluation of Commercially Available Anti\u2013Dengue Virus Immunoglobulin M Tests","volume":"15","author":"EA Hunsperger","year":"2009","journal-title":"Emerg Infect Dis"},{"issue":"3","key":"pgph.0002625.ref042","first-page":"e00711","article-title":"Performance Evaluation of a Dengue IgG Rapid Diagnostic Test Designed to Determine Dengue Serostatus as Part of Prevaccination Screening","volume":"10","author":"V Liberal","journal-title":"Microbiol Spectr"},{"issue":"6","key":"pgph.0002625.ref043","doi-asserted-by":"crossref","first-page":"e427","DOI":"10.1016\/S2666-5247(22)00033-7","article-title":"Accuracy and efficacy of pre-dengue vaccination screening for previous dengue infection with a new dengue rapid diagnostic test: a retrospective analysis of phase 3 efficacy trials","volume":"3","author":"SJ Savarino","year":"2022","journal-title":"The Lancet Microbe"},{"issue":"6","key":"pgph.0002625.ref044","doi-asserted-by":"crossref","first-page":"e0006573","DOI":"10.1371\/journal.pntd.0006573","article-title":"Accuracy of dengue clinical diagnosis with and without NS1 antigen rapid test: Comparison between human and Bayesian network model decision","volume":"12","author":"C Sa-ngamuang","year":"2018","journal-title":"PLoS Negl Trop Dis"},{"issue":"7","key":"pgph.0002625.ref045","doi-asserted-by":"crossref","first-page":"206","DOI":"10.3390\/bios11070206","article-title":"Dengue Detection: Advances in Diagnostic Tools from Conventional Technology to Point of Care","volume":"11","author":"MA Kabir","year":"2021","journal-title":"Biosensors (Basel)"},{"key":"pgph.0002625.ref046","unstructured":"Prequalified In Vitro Diagnostics | WHO\u2014Prequalification of Medical Products (IVDs, Medicines, Vaccines and Immunization Devices, Vector Control) [Internet]. [cited 2024 Jul 10]. Available from: https:\/\/extranet.who.int\/prequal\/vitro-diagnostics\/prequalified-vitro-diagnostics."},{"key":"pgph.0002625.ref047","unstructured":"CDC. Laboratory Testing Requirements for Vaccination with Dengvaxia Dengue Vaccine [Internet]. Dengue. 2024 [cited 2024 Aug 28]. Available from: https:\/\/www.cdc.gov\/dengue\/hcp\/vaccine\/testing.html."},{"key":"pgph.0002625.ref048","article-title":"The OIE ASF Reference Laboratory Network\u2019s overview of African swine fever diagnostic tests for field application","author":"K Inui","year":"2022"},{"key":"pgph.0002625.ref049","unstructured":"Terrestrial Code Online Access [Internet]. WOAH\u2014World Organisation for Animal Health. [cited 2023 Sep 11]. Available from: https:\/\/www.woah.org\/en\/what-we-do\/standards\/codes-and-manuals\/terrestrial-code-online-access\/."},{"key":"pgph.0002625.ref050","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1186\/s12917-016-0831-4","article-title":"Development of a novel lateral flow assay for detection of African swine fever in blood","volume":"12","author":"P Sastre","year":"2016","journal-title":"BMC Vet Res"},{"key":"pgph.0002625.ref051","doi-asserted-by":"crossref","first-page":"113975","DOI":"10.1016\/j.jviromet.2020.113975","article-title":"Evaluation of the diagnostic accuracy of an affordable rapid diagnostic test for African Swine Fever antigen detection in Lao People\u2019s Democratic Republic","volume":"286","author":"N Matsumoto","year":"2020","journal-title":"J Virol Methods"},{"issue":"5","key":"pgph.0002625.ref052","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1177\/1040638716654942","article-title":"Development of a duplex lateral flow assay for simultaneous detection of antibodies against African and Classical swine fever viruses","volume":"28","author":"P Sastre","year":"2016","journal-title":"J Vet Diagn Invest"},{"issue":"3","key":"pgph.0002625.ref053","doi-asserted-by":"crossref","first-page":"602","DOI":"10.7589\/2016-05-112","article-title":"Evaluation of a Commercial Field Test to Detect African Swine Fever","volume":"53","author":"S Cappai","year":"2017","journal-title":"J Wildl Dis"},{"issue":"1","key":"pgph.0002625.ref054","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1093\/clinchem\/hvab194","article-title":"\u2019Farrell BJ. Lateral Flow Assays in Infectious Disease Diagnosis","volume":"68","author":"O Boehringer HR","year":"2022","journal-title":"Clinical Chemistry"},{"issue":"11","key":"pgph.0002625.ref055","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1056\/NEJMsr2022263","article-title":"Rapid Scaling Up of Covid-19 Diagnostic Testing in the United States\u2014The NIH RADx Initiative","volume":"383","author":"BJ Tromberg","year":"2020","journal-title":"N Engl J Med"}],"container-title":["PLOS Global Public Health"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pgph.0002625","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,25]],"date-time":"2024-11-25T18:41:53Z","timestamp":1732560113000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pgph.0002625"}},"subtitle":[],"editor":[{"given":"Sadia","family":"Shakoor","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2024,11,25]]},"references-count":55,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,11,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pgph.0002625","relation":{},"ISSN":["2767-3375"],"issn-type":[{"value":"2767-3375","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,25]]}}}