{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T19:55:33Z","timestamp":1784231733509,"version":"3.55.0"},"reference-count":36,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T00:00:00Z","timestamp":1681689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001321","name":"National Research Foundation (NRF) of South Africa","doi-asserted-by":"publisher","award":["98339"],"award-info":[{"award-number":["98339"]}],"id":[{"id":"10.13039\/501100001321","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001321","name":"National Research Foundation (NRF) of South Africa","doi-asserted-by":"publisher","award":["HDTRA1-20-1-0025"],"award-info":[{"award-number":["HDTRA1-20-1-0025"]}],"id":[{"id":"10.13039\/501100001321","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Department of the Defense, Defense Threat Reduction Agency","award":["98339"],"award-info":[{"award-number":["98339"]}]},{"name":"Department of the Defense, Defense Threat Reduction Agency","award":["HDTRA1-20-1-0025"],"award-info":[{"award-number":["HDTRA1-20-1-0025"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Recent studies have indicated that bats are hosts to diverse filoviruses. Currently, no pan-filovirus molecular assays are available that have been evaluated for the detection of all mammalian filoviruses. In this study, a two-step pan-filovirus SYBR Green real-time PCR assay targeting the nucleoprotein gene was developed for filovirus surveillance in bats. Synthetic constructs were designed as representatives of nine filovirus species and used to evaluate the assay. This assay detected all synthetic constructs included with an analytical sensitivity of 3\u201331.7 copies\/reaction and was evaluated against the field collected samples. The assay\u2019s performance was similar to a previously published probe based assay for detecting Ebola- and Marburgvirus. The developed pan-filovirus SYBR Green assay will allow for more affordable and sensitive detection of mammalian filoviruses in bat samples.<\/jats:p>","DOI":"10.3390\/v15040987","type":"journal-article","created":{"date-parts":[[2023,4,18]],"date-time":"2023-04-18T02:30:03Z","timestamp":1681785003000},"page":"987","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Development of a Pan-Filoviridae SYBR Green qPCR Assay for Biosurveillance Studies in Bats"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8376-6508","authenticated-orcid":false,"given":"Jessica","family":"Coertse","sequence":"first","affiliation":[{"name":"Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, A Division of the National Health Laboratory Service, Johannesburg 2131, South Africa"},{"name":"Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9286-1040","authenticated-orcid":false,"given":"Marinda","family":"Mortlock","sequence":"additional","affiliation":[{"name":"Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antoinette","family":"Grobbelaar","sequence":"additional","affiliation":[{"name":"Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, A Division of the National Health Laboratory Service, Johannesburg 2131, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naazneen","family":"Moolla","sequence":"additional","affiliation":[{"name":"Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, A Division of the National Health Laboratory Service, Johannesburg 2131, South Africa"},{"name":"Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7550-0080","authenticated-orcid":false,"given":"Wanda","family":"Markotter","sequence":"additional","affiliation":[{"name":"Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9471-2890","authenticated-orcid":false,"given":"Jacqueline","family":"Weyer","sequence":"additional","affiliation":[{"name":"Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, A Division of the National Health Laboratory Service, Johannesburg 2131, South Africa"},{"name":"Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa"},{"name":"Department of Microbiology and Infectious Diseases, School of Pathology, University of Witwatersrand, Johannesburg 2131, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3513","DOI":"10.1007\/s00705-021-05143-6","article-title":"2021 Taxonomic Update of Phylum Negarnaviricota (Riboviria: Orthornavirae), Including the Large Orders Bunyavirales and Mononegavirales","volume":"166","author":"Kuhn","year":"2021","journal-title":"Arch. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"E6","DOI":"10.1038\/s41586-018-0310-0","article-title":"Author Correction: The Evolutionary History of Vertebrate RNA Viruses","volume":"561","author":"Shi","year":"2018","journal-title":"Nature"},{"key":"ref_3","unstructured":"Emanuel, J., Marzi, A., and Feldmann, H. (2018). Advances in Virus Research, Elsevier Inc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2341","DOI":"10.1055\/s-0028-1106144","article-title":"Zur \u00c4tiologie Einer Unbekannten, von Affen Ausgegangenen Menschlichen Infektionskrankheit","volume":"92","author":"Siegert","year":"1967","journal-title":"DMW\u2014Dtsch. Med. Wochenschr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1178122X1984992","DOI":"10.1177\/1178122X19849927","article-title":"Filovirus Disease Outbreaks: A Chronological Overview","volume":"10","author":"Languon","year":"2019","journal-title":"Virol. Res. Treat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1016\/j.jiph.2020.03.014","article-title":"Ebola Outbreak in West Africa, 2014\u20132016: Epidemic Timeline, Differential Diagnoses, Determining Factors, and Lessons for Future Response","volume":"13","author":"Kamorudeen","year":"2020","journal-title":"J. Infect. Public Health"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"321","DOI":"10.3201\/eid0204.960407","article-title":"Experimental Inoculation of Plants and Animals with Ebola Virus","volume":"2","author":"Swanepoel","year":"1996","journal-title":"Emerg. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1038\/438575a","article-title":"Fruit Bats as Reservoirs of Ebola Virus","volume":"438","author":"Leroy","year":"2005","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1847","DOI":"10.3201\/eid1312.071115","article-title":"Studies of Reservoir Hosts for Marburg Virus","volume":"13","author":"Swanepoel","year":"2007","journal-title":"Emerg. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Towner, J.S., Amman, B.R., Sealy, T.K., Carroll, S.A.R., Comer, J.A., Kemp, A., Swanepoel, R., Paddock, C.D., Balinandi, S., and Khristova, M.L. (2009). Isolation of Genetically Diverse Marburg Viruses from Egyptian Fruit Bats. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000536"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Negredo, A., Palacios, G., V\u00e1zquez-Mor\u00f3n, S., Gonz\u00e1lez, F., Dopazo, H., Molero, F., Juste, J., Quetglas, J., Savji, N., and de la Cruz Mart\u00ednez, M. (2011). Discovery of an Ebolavirus-Like Filovirus in Europe. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1002304"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41426-018-0067-4","article-title":"Re-Emergence of Lloviu Virus in Miniopterus schreibersii Bats, Hungary, 2016","volume":"7","author":"Kemenesi","year":"2018","journal-title":"Emerg. Microbes Infect."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1706","DOI":"10.1038\/s41467-022-29298-1","article-title":"Isolation of Infectious Lloviu Virus from Schreiber\u2019s Bats in Hungary","volume":"13","author":"Kemenesi","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1486","DOI":"10.1038\/s41564-018-0315-3","article-title":"Author Correction: The Discovery of Bombali Virus Adds Further Support for Bats as Hosts of Ebolaviruses","volume":"3","author":"Goldstein","year":"2018","journal-title":"Nat. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2583","DOI":"10.3201\/eid2812.220853","article-title":"Bombali ebolavirus in Mops Condylurus Bats (Molossidae), Mozambique","volume":"28","author":"Lebarbenchon","year":"2022","journal-title":"Emerg. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"955","DOI":"10.3201\/eid2505.181666","article-title":"Bombali Virus in Mops Condylurus Bat, Kenya","volume":"25","author":"Forbes","year":"2019","journal-title":"Emerg. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1038\/s41564-018-0328-y","article-title":"Characterization of a Filovirus (M\u011bngl\u00e0 Virus) from Rousettus Bats in China","volume":"4","author":"Yang","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"482","DOI":"10.3201\/eid2303.161119","article-title":"Genetically Diverse Filoviruses in Rousettus and Eonycteris Spp. Bats, China, 2009 and 2015","volume":"23","author":"Yang","year":"2017","journal-title":"Emerg. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1016\/0140-6736(90)90737-P","article-title":"Preliminary Report: Isolation of Ebola Virus from Monkeys Imported to USA","volume":"335","author":"Jahrling","year":"1990","journal-title":"Lancet"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1186\/s12985-015-0331-3","article-title":"Molecular Evidence of Ebola Reston Virus Infection in Philippine Bats","volume":"12","author":"Jayme","year":"2015","journal-title":"Virol. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1759","DOI":"10.3390\/v6041759","article-title":"Filoviruses in Bats: Current Knowledge and Future Directions","volume":"6","author":"Olival","year":"2014","journal-title":"Viruses"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.1016\/S0140-6736(95)90925-7","article-title":"Isolation and Partial Characterisation of a New Strain of Ebola Virus","volume":"345","author":"Formenty","year":"1995","journal-title":"Lancet"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jcv.2018.03.005","article-title":"The Current Landscape of Nucleic Acid Tests for Filovirus Detection","volume":"103","author":"Clark","year":"2018","journal-title":"J. Clin. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.jcv.2019.03.010","article-title":"Development, Validation and Clinical Evaluation of a Broad-Range Pan-Filovirus RT-QPCR","volume":"114","author":"Sironen","year":"2019","journal-title":"J. Clin. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Young, C.C.W., and Olival, K.J. (2016). Optimizing Viral Discovery in Bats. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0149237"},{"key":"ref_26","unstructured":"Simmons, N.B., and Cirranello, A.L. (2022, September 09). Bat Species of the World: A Taxonomic and Geographic Database. Available online: https:\/\/batnames.org\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1111\/jzo.12769","article-title":"Bat-borne Viruses in Africa: A Critical Review","volume":"311","author":"Markotter","year":"2020","journal-title":"J. Zool."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Giannini, N.P., Gunnell, G.F., Habersetzer, J., and Simmons, N.B. (2012). Evolutionary History of Bats, Cambridge University Press.","DOI":"10.1017\/CBO9781139045599"},{"key":"ref_29","first-page":"95","article-title":"BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95\/98\/NT","volume":"41","author":"Hall","year":"1999","journal-title":"Nucleic Acids Symp. Ser."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"S199","DOI":"10.1086\/520600","article-title":"Diagnostic Reverse-Transcription Polymerase Chain Reaction Kit for Filoviruses Based on the Strain Collections of All European Biosafety Level 4 Laboratories","volume":"196","author":"Panning","year":"2007","journal-title":"J. Infect. Dis."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Paweska, J.T., Jansen van Vuren, P., Masumu, J., Leman, P.A., Grobbelaar, A.A., Birkhead, M., Clift, S., Swanepoel, R., and Kemp, A. (2012). Virological and Serological Findings in Rousettus aegyptiacus Experimentally Inoculated with Vero Cells-Adapted Hogan Strain of Marburg Virus. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0045479"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"24262","DOI":"10.1038\/s41598-021-03641-w","article-title":"Seasonal Shedding Patterns of Diverse Henipavirus-Related Paramyxoviruses in Egyptian Rousette Bats","volume":"11","author":"Mortlock","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_34","unstructured":"(2023, January 10). Ebolvirus Disease Outbreaks, Available online: https:\/\/www.cdc.gov\/vhf\/ebola\/outbreaks\/index-2018.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fvhf%2Febola%2Foutbreaks%2Findex.html."},{"key":"ref_35","unstructured":"(2023, January 10). History of Marburg Virus Disease (MVD) Outbreaks, Available online: https:\/\/www.cdc.gov\/vhf\/marburg\/outbreaks\/chronology.html."},{"key":"ref_36","unstructured":"Van Cakenberghe, V., and Seamark, E.C. (2022). African Chiroptera Report 2022, AfricanBats NPC."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/4\/987\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:17:36Z","timestamp":1760123856000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/4\/987"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,17]]},"references-count":36,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,4]]}},"alternative-id":["v15040987"],"URL":"https:\/\/doi.org\/10.3390\/v15040987","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,17]]}}}