{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T05:14:37Z","timestamp":1761801277535,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:00:00Z","timestamp":1761609600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the National Science and Technology Major Project","award":["2018ZX10101003-005-010"],"award-info":[{"award-number":["2018ZX10101003-005-010"]}]},{"name":"the National Key Research and Development Program of China","award":["2021YFE0200600"],"award-info":[{"award-number":["2021YFE0200600"]}]},{"DOI":"10.13039\/501100001809","name":"the National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["21776071"],"award-info":[{"award-number":["21776071"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Human norovirus (HuNoV), particularly the GII.4 genotype, is a leading cause of acute gastroenteritis worldwide, posing a significant public health and economic burden due to its low infectious dose. To address the need for rapid and sensitive detection, we developed a colorimetric biosensor utilizing a structure-optimized aptamer and gold nanoparticles (AuNPs). Biotin-modified aptamers could protect AuNPs from aggregation in salt solution. Upon specific binding to GII.4 HuNoV virus-like particles (VLPs), this protective effect is disrupted, leading to AuNP aggregation and a measurable color shift quantified by the A620\/A520 absorbance ratio. Under optimized conditions, the assay demonstrated a linear response (y = 0.004597x + 0.3277, R2 = 0.9922) to GII.4 HuNoV VLP concentrations ranging from 0.1 to 3.0 \u03bcg\/mL, with the recovery rates between 91.74% and 106.43%. The biosensor exhibited high specificity for GII.4 HuNoV, showing minimal cross-reactivity with other common diarrheal pathogens, and achieved an exceptional detection limit of 27.2 copies\/mL in a fecal matrix. Molecular docking and point mutation confirmed the critical roles of specific nucleotide bases (T20, C22, G31, and G44) in the aptamer and the Asn55 residue in the viral capsid for binding. This work establishes a sensitive, rapid, and cost-effective aptamer-based colorimetric platform suitable for the large-scale monitoring of GII.4 HuNoV.<\/jats:p>","DOI":"10.3390\/bios15110713","type":"journal-article","created":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T05:48:46Z","timestamp":1761716926000},"page":"713","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimized Aptamer-Conjugated Gold Nanoparticles for Specific Detection of GII.4 Human Norovirus in Feces"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1303-5521","authenticated-orcid":false,"given":"Chao","family":"Cheng","sequence":"first","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaomeng","family":"Zhang","sequence":"additional","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gaoyang","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory for Advanced Materials, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minjia","family":"Sun","sequence":"additional","affiliation":[{"name":"Key Laboratory for Advanced Materials, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China"},{"name":"Zhejiang CONBA Pharmaceutical Co., Ltd., Hangzhou 310052, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5692-5959","authenticated-orcid":false,"given":"Wenjing","family":"Zheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Medical Molecular Virology of MoE & MoH, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingjing","family":"Li","sequence":"additional","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9728-8893","authenticated-orcid":false,"given":"Jing","family":"Liu","sequence":"additional","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuanyi","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Medical Molecular Virology of MoE & MoH, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Youhua","family":"Xie","sequence":"additional","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6019-0585","authenticated-orcid":false,"given":"Shouhong","family":"Xu","sequence":"additional","affiliation":[{"name":"Key Laboratory for Advanced Materials, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junqi","family":"Zhang","sequence":"additional","affiliation":[{"name":"MOE\/NHC\/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Institute of Infectious Disease and Biosecurity, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1075","DOI":"10.1128\/jvi.10.5.1075-1081.1972","article-title":"Visualization by Immune Electron-Microscopy of a 27-Nm Particle Associated with Acute Infectious Nonbacterial Gastroenteritis","volume":"10","author":"Kapikian","year":"1972","journal-title":"J. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1093\/infdis\/jit620","article-title":"Determination of the 50% human infectious dose for Norwalk virus","volume":"209","author":"Atmar","year":"2014","journal-title":"J. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.coviro.2013.12.003","article-title":"Environmental persistence and transfer of enteric viruses","volume":"4","author":"Kotwal","year":"2014","journal-title":"Curr. Opin. Virol."},{"doi-asserted-by":"crossref","unstructured":"Bartsch, S.M., Lopman, B.A., Ozawa, S., Hall, A.J., and Lee, B.Y. (2016). Global Economic Burden of Norovirus Gastroenteritis. PLoS ONE, 11.","key":"ref_4","DOI":"10.1371\/journal.pone.0151219"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1468","DOI":"10.1002\/jmv.21237","article-title":"Norwalk virus: How infectious is it?","volume":"80","author":"Teunis","year":"2008","journal-title":"J. Med. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2208","DOI":"10.1128\/JCM.00455-17","article-title":"Genetic and Epidemiologic Trends of Norovirus Outbreaks in the United States from 2013 to 2016 Demonstrated Emergence of Novel GII. 4 Recombinant Viruses","volume":"55","author":"Cannon","year":"2017","journal-title":"J. Clin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2770","DOI":"10.1128\/JCM.00654-10","article-title":"Diagnostic Accuracy and Analytical Sensitivity of IDEIA Norovirus Assay for Routine Screening of Human Norovirus","volume":"48","author":"Costantini","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1002\/cbic.201300693","article-title":"Aptamers Act as Activators for the Thrombin Mediated-Hydrolysis of Peptide Substrates","volume":"15","author":"Tan","year":"2014","journal-title":"ChemBioChem"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1126\/science.2200121","article-title":"Systematic Evolution of Ligands by Exponential Enrichment\u2014Rna Ligands to Bacteriophage-T4 DNA-Polymerase","volume":"249","author":"Tuerk","year":"1990","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1016\/j.coph.2010.06.009","article-title":"Development of aptamer therapeutics","volume":"10","author":"Bunka","year":"2010","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1007\/s00604-014-1420-5","article-title":"Visual and microplate detection of aflatoxin B2 based on NaCl-induced aggregation of aptamer-modified gold nanoparticles","volume":"182","author":"Luan","year":"2015","journal-title":"Microchim. Acta"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1007\/s40242-017-7029-6","article-title":"An Aptamer-based Colorimetric Sensor for Streptomycin and Its Application in Food Inspection","volume":"33","author":"Liu","year":"2017","journal-title":"Chem. Res. Chin. Univ."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4048","DOI":"10.1039\/C7NR00612H","article-title":"A colorimetric nanosensor based on a selective target-responsive aptamer kissing complex","volume":"9","author":"Goux","year":"2017","journal-title":"Nanoscale"},{"doi-asserted-by":"crossref","unstructured":"Hu, X.R., Chang, K.K., Wang, S., Sun, X.Q., Hu, J.D., and Jiang, M. (2018). Aptamer-functionalized AuNPs for the high- sensitivity colorimetric detection of melamine in milk samples. PLoS ONE, 13.","key":"ref_14","DOI":"10.1371\/journal.pone.0201626"},{"key":"ref_15","first-page":"1","article-title":"Selection, Identification, and Application of Aptamers against Lectin to Establish an Aptamer-AuNPs Colorimetric Method for Detection of ABL","volume":"2020","author":"Gao","year":"2020","journal-title":"J. Food Qual."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1007\/s40242-013-3039-1","article-title":"Au nanoparticle-DNAzyme dual catalyst system for sensitively colorimetric detection of thrombin","volume":"29","author":"Rao","year":"2013","journal-title":"Chem. Res. Chin. Univ."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1038\/nmat2162","article-title":"Biosensing with plasmonic nanosensors","volume":"7","author":"Anker","year":"2008","journal-title":"Nat. Mater."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/S0022-2836(84)80026-1","article-title":"Kinetics and Mechanism in the Reaction of Gene Regulatory Proteins with DNA","volume":"172","author":"Fried","year":"1984","journal-title":"J. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1006\/jmbi.1998.1738","article-title":"Calorimetric studies of E. coli SSB protein-single-stranded DNA interactions. Effects of monovalent salts on binding enthalpy","volume":"278","author":"Kozlov","year":"1998","journal-title":"J. Mol. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9037","DOI":"10.1039\/C5NR08465B","article-title":"Nanoparticles and DNA\u2014A powerful and growing functional combination in bionanotechnology","volume":"8","author":"Samanta","year":"2016","journal-title":"Nanoscale"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2297","DOI":"10.1016\/j.bios.2010.09.056","article-title":"Ultrasensitive electrochemical aptasensor for thrombin based on the amplification of aptamer-AuNPs-HRP conjugates","volume":"26","author":"Zhao","year":"2011","journal-title":"Biosens. Bioelectron."},{"doi-asserted-by":"crossref","unstructured":"Papavlassopoulos, H., Mishra, Y.K., Kaps, S., Paulowicz, I., Abdelaziz, R., Elbahri, M., Maser, E., Adelung, R., and R\u00f6hl, C. (2014). Toxicity of Functional Nano-Micro Zinc Oxide Tetrapods: Impact of Cell Culture Conditions, Cellular Age and Material Properties. PLoS ONE, 9.","key":"ref_22","DOI":"10.1371\/journal.pone.0084983"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1021\/nl0727563","article-title":"A DNA-gold nanoparticle-based colorimetric competition assay for the detection of cysteine","volume":"8","author":"Lee","year":"2008","journal-title":"Nano Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1016\/j.bios.2014.02.008","article-title":"Homogeneous assay of target molecules based on chemiluminescence resonance energy transfer (CRET) using DNAzyme-linked aptamers","volume":"58","author":"Mun","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2376","DOI":"10.1007\/s11051-014-2376-4","article-title":"Aggregation behaviour of gold nanoparticles in saline aqueous media","volume":"16","author":"Pamies","year":"2014","journal-title":"J. Nanopart. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1007\/s00604-018-2895-2","article-title":"Colorimetric determination of ofloxacin using unmodified aptamers and the aggregation of gold nanoparticles","volume":"185","author":"Zhou","year":"2018","journal-title":"Microchim. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1007\/s00604-019-3261-8","article-title":"Fluorometric determination of lipopolysaccharides via changes of the graphene oxide-enhanced fluorescence polarization caused by truncated aptamers","volume":"186","author":"Ye","year":"2019","journal-title":"Microchim. Acta"},{"doi-asserted-by":"crossref","unstructured":"Sharifi, S., Vahed, S.Z., Ahmadian, E., Dizaj, S.M., Eftekhari, A., Khalilov, R., Ahmadi, M., Hamidi-Asl, E., and Labib, M. (2020). Detection of pathogenic bacteria via nanomaterials-modified aptasensors. Biosens. Bioelectron., 150.","key":"ref_28","DOI":"10.1016\/j.bios.2019.111933"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1789","DOI":"10.2147\/IJN.S495399","article-title":"Nucleic Acid Aptamers for Human Norovirus GII.4 and GII.17 Virus-like Particles (VLPs) Exhibit Specific Binding and Inhibit VLPs from Entering Cells","volume":"20","author":"Cheng","year":"2025","journal-title":"Int. J. Nanomed."},{"key":"ref_30","first-page":"1028","article-title":"Diagnosis of Norwalk virus infection by indirect enzyme immunoassay detection of salivary antibodies to recombinant Norwalk virus antigen","volume":"11","author":"Moe","year":"2004","journal-title":"Clin. Diagn. Lab. Immunol."},{"doi-asserted-by":"crossref","unstructured":"Zhou, H.L., Chen, L.N., Wang, S.M., Tan, M., Qiu, C., Qiu, T.Y., and Wang, X.Y. (2021). Prevalence and Evolution of Noroviruses between 1966 and 2019, Implications for Vaccine Design. Pathogens., 10.","key":"ref_31","DOI":"10.3390\/pathogens10081012"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3406","DOI":"10.1093\/nar\/gkg595","article-title":"Mfold web server for nucleic acid folding and hybridization prediction","volume":"31","author":"Zuker","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/S0014-5793(04)00197-8","article-title":"Screening of functional antidotes of RNA aptamers against bovine thrombin","volume":"562","author":"Liu","year":"2004","journal-title":"FEBS Lett."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1126\/science.287.5454.820","article-title":"Biochemistry\u2014Adaptive recognition by nucleic acid aptamers","volume":"287","author":"Hermann","year":"2000","journal-title":"Science."},{"doi-asserted-by":"crossref","unstructured":"Shi, M.H., Liu, R.B., Zhang, F.Y., Chitrakar, B., and Wang, X.H. (2022). Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety. Biosensors, 12.","key":"ref_35","DOI":"10.21203\/rs.3.rs-1586215\/v1"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1038\/nchem.1352","article-title":"The gold-sulfur interface at the nanoscale","volume":"4","year":"2012","journal-title":"Nat. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1038\/s41557-019-0216-y","article-title":"Non-chemisorbed gold-sulfur binding prevails in self-assembled monolayers","volume":"11","author":"Inkpen","year":"2019","journal-title":"Nat. Chem."},{"doi-asserted-by":"crossref","unstructured":"Liu, R., Zhang, F., Sang, Y., Liu, M., Shi, M., and Wang, X. (2022). Selection and Characterization of DNA Aptamers for Constructing Aptamer-AuNPs Colorimetric Method for Detection of AFM1. Foods, 11.","key":"ref_38","DOI":"10.3390\/foods11121802"},{"doi-asserted-by":"crossref","unstructured":"Martorell, S., Santiso-Bell\u00f3n, C., Gozalbo-Rovira, R., Quintero-Campos, P., Luque, D., Maquieira, A., Rodr\u00edguez-D\u00edaz, J., and Morais, S. (2025). Synthetic biology-driven optoelectronic biosensor for rapid and highly sensitive norovirus detection in fecal samples. Biosens. Bioelectron., 288.","key":"ref_39","DOI":"10.1016\/j.bios.2025.117787"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.bios.2016.08.101","article-title":"Size-controlled preparation of peroxidase-like graphene-gold nanoparticle hybrids for the visible detection of norovirus-like particles","volume":"87","author":"Ahmed","year":"2017","journal-title":"Biosens. Bioelectron."},{"doi-asserted-by":"crossref","unstructured":"Hern\u00e1ndez, O.H., Guti\u00e9rrez-Escolano, A.L., Cancio-Lonches, C., Iturriaga, M.H., Pacheco-Aguilar, J.R., Morales-Rayas, R., and Arvizu-Medrano, S.M. (2022). Multiplex PCR method for the detection of human norovirus, Salmonella spp., Shigella spp., and shiga toxin producing Escherichia coli in blackberry, coriander, lettuce and strawberry. Food Microbiol., 102.","key":"ref_41","DOI":"10.1016\/j.fm.2021.103926"},{"doi-asserted-by":"crossref","unstructured":"Li, X.Y., Zhao, Y.Q., Gu, W.C., Qian, Y., Huang, Q., Hu, X.J., and Xing, H.B. (2024). A novel dual-mode aptasensor based colorimetry and electrochemical detection of norovirus in fecal sample. Anal. Biochem., 687.","key":"ref_42","DOI":"10.1016\/j.ab.2023.115444"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"132390","DOI":"10.1016\/j.snb.2022.132390","article-title":"Impedimetric biosensor of Norovirus with low variance using simple bioconjugation on conductive polymer-Au nanocomposite","volume":"369","author":"Nasrin","year":"2022","journal-title":"Sensor Actuators B Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e29487","DOI":"10.1002\/jmv.29487","article-title":"Rapid and sensitive lateral flow biosensor for the detection of GII human norovirus based on immunofluorescent nanomagnetic microspheres","volume":"96","author":"Gao","year":"2024","journal-title":"J. Med. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.bios.2018.09.024","article-title":"Single-step detection of norovirus tuning localized surface plasmon resonance-induced optical signal between gold nanoparticles and quantum dots","volume":"122","author":"Nasrin","year":"2018","journal-title":"Biosens. Bioelectron."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.virol.2021.09.008","article-title":"CRISPR-Cas12a combined with reverse transcription recombinase polymerase amplification for sensitive and specific detection of human norovirus genotype GII.4","volume":"564","author":"Qian","year":"2021","journal-title":"Virology"},{"key":"ref_47","first-page":"11527","article-title":"A DNA aptamer for binding and inhibition of DNA methyltransferase 1","volume":"47","author":"Wang","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"7731","DOI":"10.1021\/jacs.2c00478","article-title":"Structural Insights into the Mechanism of High-Affinity Binding of Ochratoxin A by a DNA Aptamer","volume":"144","author":"Xu","year":"2022","journal-title":"J. Am. Chem. Soc."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"107141","DOI":"10.1016\/j.jfca.2024.107141","article-title":"Ultra-sensitive label-free biosensor for doxorubicin detection by doxorubicin optimized aptamer","volume":"139","author":"Wei","year":"2025","journal-title":"J. Food Compos. Anal."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"113147","DOI":"10.1016\/j.microc.2025.113147","article-title":"Sandwich-type electrochemical aptamer-based sensor for rapid nanomolar detection of anesthetic drug procaine in biofluids","volume":"211","author":"Xie","year":"2025","journal-title":"Microchem. J."}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/11\/713\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T05:12:37Z","timestamp":1761801157000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/15\/11\/713"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,28]]},"references-count":50,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["bios15110713"],"URL":"https:\/\/doi.org\/10.3390\/bios15110713","relation":{},"ISSN":["2079-6374"],"issn-type":[{"type":"electronic","value":"2079-6374"}],"subject":[],"published":{"date-parts":[[2025,10,28]]}}}