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Analyte recognition is realized through a sandwich scheme with a capture bioreceptor layer of antibodies \u2014 covalently immobilized onto the gold nanoparticle surface in upright orientation and close-packed configuration by photochemical immobilization technique (PIT) \u2014 and a top bioreceptor layer of fluorescently labelled aptamers. Such a sandwich configuration warrants not only extremely high specificity, but also an ideal fluorophore-nanostructure distance (approximately 10\u201315\u00a0nm) for achieving strong fluorescence amplification. For a specific application, we tested the biosensor performance in a case study for the detection of malaria-related marker <jats:italic>Plasmodium falciparum<\/jats:italic> lactate dehydrogenase (<jats:italic>Pf<\/jats:italic>LDH). The proposed biosensor can specifically detect <jats:italic>Pf<\/jats:italic>LDH in spiked whole blood down to 10 pM (0.3\u00a0ng\/mL) without any sample pretreatment. The combination of simple and scalable fabrication, potentially high-throughput analysis, and excellent sensing performance provides a new approach to biosensing with significant advantages compared to conventional fluorescence immunoassays.<\/jats:p>\n                <jats:p><jats:bold>Graphical abstract<\/jats:bold><\/jats:p>","DOI":"10.1007\/s00604-021-04746-9","type":"journal-article","created":{"date-parts":[[2021,2,19]],"date-time":"2021-02-19T13:01:55Z","timestamp":1613739715000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Randomly positioned gold nanoparticles as fluorescence enhancers in apta-immunosensor for malaria test"],"prefix":"10.1007","volume":"188","author":[{"given":"Antonio","family":"Minopoli","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bartolomeo","family":"Della Ventura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raffaele","family":"Campanile","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julian A.","family":"Tanner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"Offenh\u00e4usser","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dirk","family":"Mayer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1077-8353","authenticated-orcid":false,"given":"Raffaele","family":"Velotta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,2,16]]},"reference":[{"key":"4746_CR1","doi-asserted-by":"publisher","first-page":"1801385","DOI":"10.1002\/smll.201801385","volume":"14","author":"NHT Tran","year":"2018","unstructured":"Tran NHT, Trinh KTL, Lee J-H, Yoon WJ, Ju H (2018) Reproducible enhancement of fluorescence by bimetal mediated surface plasmon coupled emission for highly sensitive quantitative diagnosis of double-stranded DNA. 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