{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:59:30Z","timestamp":1760057970822,"version":"build-2065373602"},"reference-count":42,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,3,5]],"date-time":"2025-03-05T00:00:00Z","timestamp":1741132800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The University of Calgary Cumming School of Medicine Graduate Student Scholarship","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Eyes High Doctoral Recruitment Scholarship","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Canadian Institute of Health Research Canada CGS-M","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"CGS-D","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"The Canadian Network on Hepatitis C (CanHepC) Masters, Doctoral, and TRR179 Fellowships","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Michael Smith Foreign Study Supplement","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Canadian Institutes of Health Research","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Alberta Innovates Summer Research Studentships","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Alberta Innovates","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Canada Research Chair","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"Canada Foundation for Innovation","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"NSERC RTI","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]},{"name":"University Health Foundation Support","award":["NPC-178912"],"award-info":[{"award-number":["NPC-178912"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Globally, an estimated 254 million people are living with chronic hepatitis B virus (HBV) infection, yet only 10.5% have been diagnosed, underscoring the urgent need to expand testing to meet the World Health Organization\u2019s HBV elimination targets by 2030. Many HBV diagnostic tests remain expensive and inaccessible in resource-limited settings. In this study, we demonstrate how individually sourced, commercially available reagents can be used to develop cost-effective in-house assays for total DNA isolation, HBV viral load quantification by (q)PCR, and qHBsAg and qHBeAg measurement using sandwich ELISA. These assays were validated using known HBV-positive and HBV-negative plasma samples (genotypes A\u2013F) and HepAD38 cells treated with tenofovir disoproxil fumarate (TDF). DNA isolation using a commercial column-based kit was compared to a high-throughput, column-free method, allowing for HBV quantification from 50 \u00b5L of plasma with lower limits of detection (LLOD) of 1.8 \u00d7 103 and 1.8 \u00d7 104 HBV DNA copies IU\/mL, respectively. Both commercial and in-house DNA isolation methods yielded comparable half-maximal effective concentration (EC50) values in TDF-treated HepAD38 cells. Additionally, in-house sandwich ELISA assays were developed for quantitative HBsAg and HBeAg detection, with LLOD values of 0.78 IU\/mL and 0.38 PEI U\/mL (Paul Ehrlich Institute), respectively. The in-house reagents for DNA isolation, molecular testing, and serological detection of HBV were estimated to be at least 10 times more cost-effective than commercially available kits, highlighting their potential for broader application in resource-limited regions.<\/jats:p>","DOI":"10.3390\/pathogens14030258","type":"journal-article","created":{"date-parts":[[2025,3,5]],"date-time":"2025-03-05T11:20:57Z","timestamp":1741173657000},"page":"258","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2750-8675","authenticated-orcid":false,"given":"Simmone","family":"D\u2019souza","sequence":"first","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute, University of Lethbridge, Lethbridge, AB T1K 3M4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Layla","family":"Al-Yasiri","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annie","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dan T.","family":"Boghici","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5148-6229","authenticated-orcid":false,"given":"Guido","family":"van Marle","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer A.","family":"Corcoran","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0627-2923","authenticated-orcid":false,"given":"Trushar R.","family":"Patel","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute, University of Lethbridge, Lethbridge, AB T1K 3M4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1472-0901","authenticated-orcid":false,"given":"Carla S.","family":"Coffin","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"},{"name":"Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 2T8, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,5]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2025, January 16). 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