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In addition to enzyme immunoassays (EIAs), rapid diagnostic tests (RDTs) are available for the detection of HBsAg in resource-poor settings. However, the available RDTs have inadequate sensitivity and therefore are not suitable for diagnosis of patients with low levels of HBsAg and for blood screening. To provide a high-sensitivity RDT, we developed a lateral flow immunoassay (LFIA) for HBsAg utilizing upconverting nanoparticle (UCNP) reporter. The UCNP-LFIA can use whole blood, serum, or plasma and the results can be read in 30\u00a0min using a reader device. When compared with a commercial conventional visually read LFIA, the developed UCNP-LFIA had a Limit of Detection (LoD) of 0.1\u00a0IU HBsAg\/ml in spiked serum, whereas the LoD of the conventional LFIA was 3.2\u00a0IU HBsAg\/ml. The developed UCNP-LFIA fulfills the WHO criterion for blood screening (LoD \u2264\u20090.13\u00a0IU HBsAg\/ml) in terms of LoD. The UCNP-LFIA and conventional LFIA were evaluated with well-characterized sample panels. The UCNP-LFIA detected 20\/24 HBsAg-positive samples within the HBsAg Performance Panel and 8\/10 samples within the Mixed Titer Performance Panel, whereas the conventional LFIA detected 8\/24 and 4\/10 samples in these panels, respectively. The performance of the assays was further evaluated with HBsAg-positive (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u2009108) and HBsAg-negative (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u2009315) patient samples. In comparison with a central laboratory test, UCNP-LFIA showed 95.4% (95% CI: 89.5\u201398.5%) sensitivity whereas sensitivity of the conventional LFIA was 87.7% (95%CI: 79.9\u201393.3%).\n                  <\/jats:p>","DOI":"10.1007\/s00216-020-03055-z","type":"journal-article","created":{"date-parts":[[2020,11,23]],"date-time":"2020-11-23T17:03:01Z","timestamp":1606150981000},"page":"967-978","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":58,"title":["Upconverting nanoparticle reporter\u2013based highly sensitive rapid lateral flow immunoassay for hepatitis B virus surface antigen"],"prefix":"10.1007","volume":"413","author":[{"given":"Iida","family":"Martiskainen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheikh M.","family":"Talha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karoliina","family":"Vuorenp\u00e4\u00e4","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teppo","family":"Salminen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Etvi","family":"Juntunen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Souvick","family":"Chattopadhyay","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dinesh","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tytti","family":"Vuorinen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Pettersson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Navin","family":"Khanna","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1669-4310","authenticated-orcid":false,"given":"Gaurav","family":"Batra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,11,23]]},"reference":[{"key":"3055_CR1","unstructured":"Global Hepatitis Report 2017. 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Ltd. which is a joint assignee for a patent on a battery-powered UCNP reader device. The other authors declare that they have no conflict of interest.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"Not applicable.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to participate"}},{"value":"Not applicable.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"Not applicable.","order":6,"name":"Ethics","group":{"name":"EthicsHeading","label":"Code availability"}}]}}