Optimization of a rapid and sensitive nucleic acid lateral flow biosensor for hepatitis B virus detection

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dc.authoridYeşilkır Baydar, Serap/0000-0001-6311-4302
dc.authoridYesilkir Baydar, Serap/0000-0001-6311-4302
dc.authoridHusseini, Abbas Ali/0000-0002-8861-7106
dc.contributor.authorHusseini, Abbas Ali
dc.contributor.authorBaydar, Serap Yesilkir
dc.date.accessioned2024-09-11T19:50:21Z
dc.date.available2024-09-11T19:50:21Z
dc.date.issued2023
dc.departmentİstanbul Gelişim Üniversitesien_US
dc.description.abstractBackground and objectiveThe utilization of direct amplification of nucleic acid from lysate has attracted interest in the advancement of straightforward and economical point-of-care assays. Consequently, this study primarily focuses on the development of a rapid, precise, and cost-effective lateral flow biosensor for the convenient detection of HBV nucleic acid at the point-of-care. Furthermore, the study evaluates the effectiveness of the direct amplification method in comparison to purified nucleic acid samples within the context of LAMP-LF biosensing approaches.MethodsThe experiments conducted in this study utilized clinical serum samples that were confirmed as HBV-positive through real-time PCR assays. Sample preparation involved employing spin column nucleic acid purification and serum heat treatment. To amplify a 250 bp fragment of the HBV polymerase gene, three pairs of specific LAMP primers were utilized, which were biotin-labeled and FITC-labeled for detection purposes. Various incubation temperatures (ranging from 64 to 68 & DEG;C) and durations (30 min, 45 min, and 1 h) were investigated to determine the optimal conditions for the LAMP assay. The results were subsequently assessed through fluorometric analysis, white turbidity measurements, and lateral flow assay. Milenia HybriDetect1 strips, designed for immediate use, were employed to visualize the LAMP amplicons. Furthermore, the performance of the lateral flow biosensor was evaluated using 10-fold serial dilutions of a secondary standard containing a viral load of 10(8) IU/ml.ResultsThe optimization of the LAMP reaction was achieved at a temperature of 67 & DEG;C, resulting in significant turbidity after a 30-minute incubation period. When the spin column purification method was employed, varying test bands were observed for templates ranging from 10(8) IU/ml to 10(1) IU/ml viral load. However, when serum samples underwent heat treatment and the resulting supernatant was directly used for LAMP, the lateral flow assay was capable of detecting a minimum viral load of 10(3) IU/ml.ConclusionIn resource-limited settings, the LAMP-LF assay presents a promising solution for HBV testing. However, it is important to note that direct amplification without DNA purification may diminish the performance of the approach.en_US
dc.description.sponsorshipIstanbul Gelisim University Office of Scientific Research Projects [KAP-050421-AHH]en_US
dc.description.sponsorshipThis study was supported by Istanbul Gelisim University Office of Scientific Research Projects, Grant No.: KAP-050421-AHH.en_US
dc.identifier.doi10.1007/s11033-023-08730-9
dc.identifier.endpage8336en_US
dc.identifier.issn0301-4851
dc.identifier.issn1573-4978
dc.identifier.issue10en_US
dc.identifier.pmid37592176en_US
dc.identifier.scopus2-s2.0-85168099407en_US
dc.identifier.startpage8329en_US
dc.identifier.urihttps://doi.org/10.1007/s11033-023-08730-9
dc.identifier.urihttps://hdl.handle.net/11363/7616
dc.identifier.volume50en_US
dc.identifier.wosWOS:001051788000003en_US
dc.identifier.wosqualityQ3en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.language.isoenen_US
dc.publisherSpringeren_US
dc.relation.ispartofMolecular Biology Reportsen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.snmz20240903_Gen_US
dc.subjectHepatitis B virusen_US
dc.subjectPoint-of-careen_US
dc.subjectLoop-mediated isothermal amplificationen_US
dc.subjectLateral-flow assayen_US
dc.titleOptimization of a rapid and sensitive nucleic acid lateral flow biosensor for hepatitis B virus detectionen_US
dc.typeArticleen_US

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