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      Upconverting nanoparticle reporter–based highly sensitive rapid lateral flow immunoassay for hepatitis B virus surface antigen

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          Abstract

          Detection of hepatitis B Virus surface antigen (HBsAg) is an established method for diagnosing both acute and chronic hepatitis B virus (HBV) infection. 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 min 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 IU HBsAg/ml in spiked serum, whereas the LoD of the conventional LFIA was 3.2 IU HBsAg/ml. The developed UCNP-LFIA fulfills the WHO criterion for blood screening (LoD ≤ 0.13 IU 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 = 108) and HBsAg-negative ( n = 315) patient samples. In comparison with a central laboratory test, UCNP-LFIA showed 95.4% (95% CI: 89.5–98.5%) sensitivity whereas sensitivity of the conventional LFIA was 87.7% (95%CI: 79.9–93.3%).

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s00216-020-03055-z.

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          Most cited references31

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          Upconverting nanoparticles.

          Upconversion (UC) refers to nonlinear optical processes in which the sequential absorption of two or more photons leads to the emission of light at shorter wavelength than the excitation wavelength (anti-Stokes type emission). In contrast to other emission processes based on multiphoton absorption, upconversion can be efficiently excited even at low excitation densities. The most efficient UC mechanisms are present in solid-state materials doped with rare-earth ions. The development of nanocrystal research has evoked increasing interest in the development of synthesis routes which allow the synthesis of highly efficient, small UC particles with narrow size distribution able to form transparent solutions in a wide range of solvents. Meanwhile, high-quality UC nanocrystals can be routinely synthesized and their solubility, particle size, crystallographic phase, optical properties and shape can be controlled. In recent years, these particles have been discussed as promising alternatives to organic fluorophosphors and quantum dots in the field of medical imaging. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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            Hepatitis B virus infection.

            Hepatitis B virus infection is a major public health problem worldwide; roughly 30% of the world's population show serological evidence of current or past infection. Hepatitis B virus is a partly double-stranded DNA virus with several serological markers: HBsAg and anti-HBs, HBeAg and anti-HBe, and anti-HBc IgM and IgG. It is transmitted through contact with infected blood and semen. A safe and effective vaccine has been available since 1981, and, although variable, the implementation of universal vaccination in infants has resulted in a sharp decline in prevalence. Hepatitis B virus is not cytopathic; both liver damage and viral control--and therefore clinical outcome--depend on the complex interplay between virus replication and host immune response. Overall, as much as 40% of men and 15% of women with perinatally acquired hepatitis B virus infection will die of liver cirrhosis or hepatocellular carcinoma. In addition to decreasing hepatic inflammation, long-term antiviral treatment can reverse cirrhosis and reduce hepatocellular carcinoma. Development of new therapies that can improve HBsAg clearance and virological cure is warranted.
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              Chronic Hepatitis B Infection

              More than 240 million individuals worldwide are infected with chronic hepatitis B virus (HBV). Among individuals with chronic HBV infection who are untreated, 15% to 40% progress to cirrhosis, which may lead to liver failure and liver cancer.
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                Author and article information

                Contributors
                gaurav.batra@thsti.res.in
                Journal
                Anal Bioanal Chem
                Anal Bioanal Chem
                Analytical and Bioanalytical Chemistry
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                1618-2642
                1618-2650
                23 November 2020
                23 November 2020
                2021
                : 413
                : 4
                : 967-978
                Affiliations
                [1 ]GRID grid.1374.1, ISNI 0000 0001 2097 1371, Department of Biotechnology, , University of Turku, ; 20014 Turku, Finland
                [2 ]GRID grid.464764.3, ISNI 0000 0004 1763 2258, Translational Health Science and Technology Institute, , NCR Biotech Science Cluster, ; Faridabad, 121001 India
                [3 ]Arrow weighing systems Pvt Ltd (unit Designinnova), New Delhi, 110028 India
                [4 ]GRID grid.1374.1, ISNI 0000 0001 2097 1371, Department of Virology and Clinical Microbiology, , University of Turku, ; 20520 Turku, Finland
                [5 ]GRID grid.425195.e, ISNI 0000 0004 0498 7682, International Centre for Genetic Engineering & Biotechnology, ; New Delhi, 110067 India
                Author information
                http://orcid.org/0000-0003-1669-4310
                Article
                3055
                10.1007/s00216-020-03055-z
                7813740
                33230700
                94676598-710b-446d-a4a3-66eb11e2b626
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 22 August 2020
                : 22 October 2020
                : 9 November 2020
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100004440, Wellcome Trust;
                Award ID: 106295/A/14/Z
                Categories
                Research Paper
                Custom metadata
                © Springer-Verlag GmbH Germany, part of Springer Nature 2021

                Analytical chemistry
                hbsag,hbv diagnostics,upconverting nanophosphors,lateral flow,point-of-care test

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