{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T00:53:21Z","timestamp":1781225601638,"version":"3.54.1"},"reference-count":54,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,8,29]],"date-time":"2024-08-29T00:00:00Z","timestamp":1724889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Infectious diseases caused by fungal sources are of great interest owing to their increasing prevalence. Invasive fungal infections, including invasive pulmonary aspergillosis caused by Aspergillus fumigatus, and Pneumocystis pneumonia caused by Pneumocystis jirovecii, are significant causes of morbidity and mortality among immunocompromised patients. The accurate and timely detection of these pathogens in this high-risk population is crucial for effective patient management. We developed a multiplex real-time polymerase chain reaction (PCR) assay, RF2 mRT-PCR, specifically designed to detect two respiratory fungi, P. jirovecii and A. fumigatus, and evaluated its performance in specimens of patients with lower respiratory tract infection. The performance was evaluated using 731 clinical samples, 55 reference species, and one synthetic DNA. The reproducibility test yielded a probit curve with a lower limit of detection of 19.82 copies\/reaction for P. jirovecii and 64.20 copies\/reaction for A. fumigatus. The RF2 mRT-PCR assay did not cross-react with non-A. fumigatus\u00a0Aspergillus species or other common bacterial and viral species, and showed 100% in vitro sensitivity and specificity with reference assays. Additionally, it simultaneously detected A. fumigatus and P. jirovecii in co-infected samples. Therefore, the RF2 mRT-PCR assay is an efficient and reliable tool for in vitro diagnosis of A. fumigatus and P. jirovecii pulmonary infections.<\/jats:p>","DOI":"10.3390\/jof10090619","type":"journal-article","created":{"date-parts":[[2024,8,29]],"date-time":"2024-08-29T11:12:13Z","timestamp":1724929933000},"page":"619","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development of a Multiplex Real-Time PCR Assay for the Simultaneous Detection of Two Fungal Pathogens Causing Pneumonia"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0115-5170","authenticated-orcid":false,"given":"Ho-Jae","family":"Lim","sequence":"first","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3098-4710","authenticated-orcid":false,"given":"Seojin","family":"Ahn","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jee-Hyun","family":"No","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min-Young","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min-Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong-Hak","family":"Sohn","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7762-9593","authenticated-orcid":false,"given":"Kwang-Soo","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Graduate School, Daejeon University, Daejeon 34520, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8789-9829","authenticated-orcid":false,"given":"Jung-Eun","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Integrative Biological Sciences & BK21 FOUR Educational Research Group for Age-Associated Disorder Control Technology, Chosun University, Gwangju 61452, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6153-9646","authenticated-orcid":false,"given":"Yong-Jin","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1038\/s41564-022-01112-0","article-title":"Evolution of the human pathogenic lifestyle in fungi","volume":"7","author":"Rokas","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_2","unstructured":"World Health Organization (WHO) Report (2024, June 04). WHO Fungal Priority Pathogens List to Guide Research, Development and Public Health Action. Available online: https:\/\/www.who.int\/publications\/i\/item\/9789240060241."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e428","DOI":"10.1016\/S1473-3099(23)00692-8","article-title":"Global incidence and mortality of severe fungal disease","volume":"24","author":"Denning","year":"2024","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1183\/09059180.00001011","article-title":"Pulmonary aspergillosis: A clinical review","volume":"20","author":"Kousha","year":"2011","journal-title":"Eur. Respir. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/s12281-010-0029-3","article-title":"Pneumocystis Jirovecii Pneumonia: Current Knowledge and Outstanding Public Health Issues","volume":"4","author":"Harris","year":"2010","journal-title":"Curr. Fungal Infect. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Li, Z., Lu, G., and Meng, G. (2019). Pathogenic Fungal Infection in the Lung. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.01524"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Caceres, D.H., Rodriguez-Barradas, M.C., Whitaker, M., Jackson, B.R., Kim, L., Surie, D., Cikesh, B., Lindsley, M.D., McCotter, O.Z., and Berkow, E.L. (2023). Fungal Pathogens as Causes of Acute Respiratory Illness in Hospitalized Veterans: Frequency of Fungal Positive Test Results Using Rapid Immunodiagnostic Assays. J. Fungi, 9.","DOI":"10.3390\/jof9040456"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1093\/cid\/ciy776","article-title":"Estimation of Direct Healthcare Costs of Fungal Diseases in the United States","volume":"68","author":"Benedict","year":"2019","journal-title":"Clin. Infect. Dis."},{"key":"ref_9","unstructured":"US Centers for Disease Control and Prevention Explore Topics (2024, April 24). Clinical Overview of Aspergillosis, Available online: https:\/\/www.cdc.gov\/aspergillosis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"220114","DOI":"10.1183\/16000617.0114-2022","article-title":"Pulmonary aspergillosis: Diagnosis and treatment","volume":"31","author":"Lamoth","year":"2022","journal-title":"Eur. Respir. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Stemler, J., Tobben, C., Lass-Florl, C., Steinmann, J., Ackermann, K., Rath, P.M., Simon, M., Cornely, O.A., and Koehler, P. (2023). Diagnosis and Treatment of Invasive Aspergillosis Caused by Non-fumigatus Aspergillus spp. J. Fungi, 9.","DOI":"10.3390\/jof9040500"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1093\/mmy\/myaa024","article-title":"Diagnosing Pneumocystis jirovecii pneumonia: A review of current methods and novel approaches","volume":"58","author":"Bateman","year":"2020","journal-title":"Med. Mycol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1111\/imj.15591","article-title":"Consensus guidelines for the diagnosis and management of invasive aspergillosis, 2021","volume":"51","author":"Douglas","year":"2021","journal-title":"Intern. Med. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1016\/j.cmi.2018.01.002","article-title":"Diagnosis and management of Aspergillus diseases: Executive summary of the 2017 ESCMID-ECMM-ERS guideline","volume":"24","author":"Ullmann","year":"2018","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1007\/s00134-019-05906-5","article-title":"Diagnosis of severe respiratory infections in immunocompromised patients","volume":"46","author":"Azoulay","year":"2020","journal-title":"Intensive Care Med."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Chen, Y., Ai, L., Zhou, Y., Zhao, Y., Huang, J., Tang, W., and Liang, Y. (2021). Rapid and precise diagnosis of pneumonia coinfected by Pneumocystis jirovecii and Aspergillus fumigatus assisted by next-generation sequencing in a patient with systemic lupus erythematosus: A case report. Ann. Clin. Microbiol. Antimicrob., 20.","DOI":"10.1186\/s12941-021-00448-5"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"101330","DOI":"10.1016\/j.mycmed.2022.101330","article-title":"Clinical characteristics and risk factors of in-hospital mortality in patients coinfected with Pneumocystis jirovecii and Aspergillus","volume":"33","author":"Zhong","year":"2023","journal-title":"J. Mycol. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1111\/myc.12642","article-title":"Pulmonary co-infections by Pneumocystis jirovecii and Aspergillus fumigatus in non-HIV patients: A report of two cases and literature review","volume":"60","author":"Markantonatou","year":"2017","journal-title":"Mycoses"},{"key":"ref_19","first-page":"235","article-title":"Coinfection of invasive pulmonary aspergillosis and pneumocystis jiroveci pneumonia in a non-HIV patient","volume":"61","author":"Kaira","year":"2007","journal-title":"Acta Med. Okayama"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1016\/j.jmii.2018.11.001","article-title":"Coinfection of pneumocystis jiroveci pneumonia and pulmonary aspergillosis in a non-HIV-infected patient","volume":"51","author":"Lee","year":"2018","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1016\/j.sjbs.2020.04.032","article-title":"Pneumocystis Jirovecii detection and comparison of multiple diagnostic methods with quantitative real-time PCR in patients with respiratory symptoms","volume":"27","author":"Alshahrani","year":"2020","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Scharmann, U., Kirchhoff, L., Hain, A., Buer, J., Koldehoff, M., Steinmann, J., and Rath, P.M. (2021). Evaluation of Three Commercial PCR Assays for the Detection of Azole-Resistant Aspergillus fumigatus from Respiratory Samples of Immunocompromised Patients. J. Fungi, 7.","DOI":"10.3390\/jof7020132"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1128\/JCM.00667-15","article-title":"Analytical and Clinical Evaluation of the PathoNostics AsperGenius Assay for Detection of Invasive Aspergillosis and Resistance to Azole Antifungal Drugs during Testing of Serum Samples","volume":"53","author":"White","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.diagmicrobio.2016.10.005","article-title":"Evaluation of a new commercial real-time PCR assay for diagnosis of Pneumocystis jirovecii pneumonia and identification of dihydropteroate synthase (DHPS) mutations","volume":"87","author":"Montesinos","year":"2017","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"myab080","DOI":"10.1093\/mmy\/myab080","article-title":"Comparison of the PneumoGenius\u00ae and RealStar\u00ae Pneumocystis jirovecii PCR CE-IVD assays with a lab developed test for the detection of Pneumocystis jirovecii","volume":"60","author":"Nijhuis","year":"2022","journal-title":"Med. Mycol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1559","DOI":"10.1128\/AEM.02383-08","article-title":"Sequencing and analysis of fungal rRNA operons for development of broad-range fungal PCR assays","volume":"75","author":"Khot","year":"2009","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3210","DOI":"10.1128\/JCM.01032-17","article-title":"Molecular Diagnosis of Invasive Aspergillosis and Detection of Azole Resistance by a Newly Commercialized PCR Kit","volume":"55","author":"Dannaoui","year":"2017","journal-title":"J. Clin. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.jmoldx.2017.12.005","article-title":"Evaluation of Two Commercial Real-Time PCR Kits for Aspergillus DNA Detection in Bronchoalveolar Lavage Fluid in Patients with Invasive Pulmonary Aspergillosis","volume":"20","author":"Denis","year":"2018","journal-title":"J. Mol. Diagn."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Ruiz-Ruiz, S., Ponce, C.A., Pesantes, N., Bustamante, R., Gatti, G., San Martin, V., Gutierrez, M., Borquez, P., Vargas, S.L., and Magne, F. (2021). A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.787554"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1016\/j.jmii.2019.08.012","article-title":"Diagnosis of Pneumocystis pneumonia by real-time PCR in patients with various underlying diseases","volume":"53","author":"Yang","year":"2020","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e40","DOI":"10.1093\/nar\/gkm051","article-title":"Dual priming oligonucleotide system for the multiplex detection of respiratory viruses and SNP genotyping of CYP2C19 gene","volume":"35","author":"Chun","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4943","DOI":"10.1007\/s00253-019-09835-7","article-title":"A TaqMan-probe-based multiplex real-time RT-qPCR for simultaneous detection of porcine enteric coronaviruses","volume":"103","author":"Huang","year":"2019","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"100115","DOI":"10.1016\/j.simyco.2021.100115","article-title":"Aspergillus fumigatus and aspergillosis: From basics to clinics","volume":"100","author":"Arastehfar","year":"2021","journal-title":"Stud. Mycol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"a019786","DOI":"10.1101\/cshperspect.a019786","article-title":"Aspergillus fumigatus and related species","volume":"5","author":"Sugui","year":"2014","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lim, H.J., Kang, E.R., Park, M.Y., Kim, B.K., Kim, M.J., Jung, S., Roh, K.H., Sung, N., Yang, J.H., and Lee, M.W. (2021). Development of a multiplex real-time PCR assay for the simultaneous detection of four bacterial pathogens causing pneumonia. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0253402"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Lim, H.J., Jung, H.S., Park, M.Y., Baek, Y.H., Kannappan, B., Park, J.Y., Yang, J.H., Seol, J.H., Lee, M.W., and Jung, S.K. (2022). Evaluation of Three Automated Extraction Systems for the Detection of SARS-CoV-2 from Clinical Respiratory Specimens. Life, 12.","DOI":"10.3390\/life12010068"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1080\/13693780310001656786","article-title":"PCR identification system for the genus Aspergillus and three major pathogenic species: Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger","volume":"42","author":"Sugita","year":"2004","journal-title":"Med. Mycol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4150","DOI":"10.1128\/JCM.00570-11","article-title":"Molecular detection and species-specific identification of medically important Aspergillus species by real-time PCR in experimental invasive pulmonary aspergillosis","volume":"49","author":"Walsh","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Lim, H.J., Park, M.Y., Jung, H.S., Kwon, Y., Kim, I., Kim, D.K., Yu, N., Sung, N., Lee, S.H., and Park, J.E. (2021). Development of an efficient Sanger sequencing-based assay for detecting SARS-CoV-2 spike mutations. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0260850"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.watres.2016.03.026","article-title":"Determining the 95% limit of detection for waterborne pathogen analyses from primary concentration to qPCR","volume":"96","author":"Stokdyk","year":"2016","journal-title":"Water Res."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Henke, E., Zoch, M., Peng, Y., Reinecke, I., Sedlmayr, M., and Bathelt, F. (2024). Conceptual design of a generic data harmonization process for OMOP common data model. BMC Med. Inform. Decis. Mak., 24.","DOI":"10.1186\/s12911-024-02458-7"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.idh.2019.01.003","article-title":"Clinician perceptions of respiratory infection risk; a rationale for research into mask use in routine practice","volume":"24","author":"Barratt","year":"2019","journal-title":"Infect. Dis. Health"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"17","DOI":"10.2353\/jmoldx.2010.090173","article-title":"Molecular diagnostics for invasive fungal infections: A call for refinement and implementation","volume":"12","author":"Procop","year":"2010","journal-title":"J. Mol. Diagn."},{"key":"ref_44","first-page":"e44356","article-title":"Challenges in the Management of Invasive Fungal Infections in the Middle East: Expert Opinion to Optimize Management Using a Multidisciplinary Approach","volume":"15","author":"AlMaghrabi","year":"2023","journal-title":"Cureus"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"6009","DOI":"10.2147\/IDR.S379043","article-title":"Evaluation of the Performance of a Multiplex Real-Time PCR Assay for the Identification of Aspergillus, Cryptococcus neoformans, and Pneumocystis jirovecii Simultaneously from Sputum in Multicenter","volume":"15","author":"Liu","year":"2022","journal-title":"Infect. Drug Resist."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/j.rcl.2022.01.006","article-title":"Invasive Fungal Pneumonia in Immunocompromised Patients","volume":"60","author":"Godoy","year":"2022","journal-title":"Radiol. Clin. N. Am."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e2019074","DOI":"10.1590\/1981-6723.07419","article-title":"Occurrence of filamentous fungi isolated from matured blue cheese","volume":"23","author":"Ribeiro","year":"2020","journal-title":"Braz. J. Food Technol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1111\/j.1469-0691.2011.03760.x","article-title":"Accuracy of beta-D-glucan for the diagnosis of Pneumocystis jirovecii pneumonia: A meta-analysis","volume":"19","author":"Karageorgopoulos","year":"2013","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e921188","DOI":"10.12659\/AJCR.921188","article-title":"Utility of Intraocular Fluid beta-D-glucan Testing in Fungal Endophthalmitis: A Series of 5 Cases","volume":"21","author":"Chen","year":"2020","journal-title":"Am. J. Case Rep."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Egger, M., Jenks, J.D., Hoenigl, M., and Prattes, J. (2020). Blood Aspergillus PCR: The Good, the Bad, and the Ugly. J. Fungi, 6.","DOI":"10.3390\/jof6010018"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1093\/cid\/ciab155","article-title":"Progress Report: Next-Generation Sequencing, Multiplex Polymerase Chain Reaction, and Broad-Range Molecular Assays as Diagnostic Tools for Fever of Unknown Origin Investigations in Adults","volume":"74","author":"Wright","year":"2022","journal-title":"Clin. Infect. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"391","DOI":"10.3343\/alm.2022.42.4.391","article-title":"Update of Guidelines for Laboratory Diagnosis of COVID-19 in Korea","volume":"42","author":"Hong","year":"2022","journal-title":"Ann. Lab. Med."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1038\/s12276-020-0452-7","article-title":"Optimization of primer sets and detection protocols for SARS-CoV-2 of coronavirus disease 2019 (COVID-19) using PCR and real-time PCR","volume":"52","author":"Park","year":"2020","journal-title":"Exp. Mol. Med."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1016\/j.pathol.2021.03.006","article-title":"Quantitative Pneumocystis jirovecii real-time PCR to differentiate disease from colonisation","volume":"53","author":"Tan","year":"2021","journal-title":"Pathology"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/9\/619\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:45:16Z","timestamp":1760111116000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/9\/619"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,29]]},"references-count":54,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2024,9]]}},"alternative-id":["jof10090619"],"URL":"https:\/\/doi.org\/10.3390\/jof10090619","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,29]]}}}