{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T09:06:09Z","timestamp":1789031169638,"version":"build-2803163510"},"reference-count":27,"publisher":"Walter de Gruyter GmbH","issue":"3","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec id=\"j_almed-2025-0197_abs_001\">\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>Rabies, a neglected zoonotic encephalitis caused by lyssaviruses, claims around 59,000 lives annually. PCR-based methods are pivotal for laboratory confirmation of rabies in humans and animals. The LN34\u00a0pan-lyssavirus real time PCR assay has demonstrated good diagnostic performance in detecting rabies viral RNA in extensive evaluations with animal brain tissues. However, its application has not been adequately evaluated on human samples.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_almed-2025-0197_abs_002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>This study assessed the utility of the LN34 assay on human brain tissues, and antemortem samples such as cerebrospinal fluid, saliva and nuchal skin. Furthermore, this assay was also evaluated using postmortem dog saliva samples, in addition to animal brain samples.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_almed-2025-0197_abs_003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Compared to the gold-standard fluorescent antibody test (FAT), the LN34 assay showed 100\u202f% sensitivity and specificity for both human and animal brain samples. For human antemortem samples, there was perfect agreement between the LN34 assay and routine real-time RT-PCR for detection of rabies virus (RABV). Similarly, there was complete agreement between the two assays when testing dog saliva samples. The sensitivity and specificity of saliva LN34 assay in laboratory-confirmed rabid dogs was 50 and 100\u202f% respectively.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_almed-2025-0197_abs_004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The LN34 assay is a suitable alternative for diagnosis and surveillance of rabies in humans and animals in an area endemic for rabies lyssavirus.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1515\/almed-2025-0197","type":"journal-article","created":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T09:55:57Z","timestamp":1783418157000},"page":"222-228","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of LN34\u00a0real-time PCR assay for detection of rabies virus in human and animal specimens"],"prefix":"10.1515","volume":"7","author":[{"given":"Lonika","family":"Lodha","sequence":"first","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manoor Ananda","family":"Ashwini","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sathya Priya","family":"Manuel","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arpita","family":"Maladakar","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shubhangi","family":"Chandel","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Preeti Soundarya","family":"Bhalke","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shrinivas","family":"Navya","sequence":"additional","affiliation":[{"name":"Department of Microbiology Veterinary College , KVAFSU-CVA Rabies Diagnostic Laboratory, WOAH Reference Laboratory for Rabies, KVAFSU , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuntanahalli Narayanaswamy","family":"Krupa","sequence":"additional","affiliation":[{"name":"Department of Microbiology Veterinary College , KVAFSU-CVA Rabies Diagnostic Laboratory, WOAH Reference Laboratory for Rabies, KVAFSU , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shrikrishna","family":"Isloor","sequence":"additional","affiliation":[{"name":"Department of Microbiology Veterinary College , KVAFSU-CVA Rabies Diagnostic Laboratory, WOAH Reference Laboratory for Rabies, KVAFSU , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Balaji","family":"Chandrashekar","sequence":"additional","affiliation":[{"name":"Mission Rabies-WVS , Goa , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anita","family":"Mahadevan","sequence":"additional","affiliation":[{"name":"Department of Neuropathology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6039-1187","authenticated-orcid":false,"given":"Reeta S.","family":"Mani","sequence":"additional","affiliation":[{"name":"Department of Neurovirology , National Institute of Mental Health and Neurosciences (NIMHANS) , Bengaluru , India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"374","published-online":{"date-parts":[[2026,7,7]]},"reference":[{"key":"2026091008070976178_j_almed-2025-0197_ref_001","doi-asserted-by":"crossref","unstructured":"Walker, PJ, Freitas-Ast\u00faa, J, Bejerman, N, Blasdell, KR, Breyta, R, Dietzgen, RG, et al.. ICTV virus taxonomy profile: rhabdoviridae 2022. J\u00a0Gen Virol 2022;103:001689. https:\/\/doi.org\/10.1099\/jgv.0.001689.","DOI":"10.1099\/jgv.0.001689"},{"key":"2026091008070976178_j_almed-2025-0197_ref_002","unstructured":"World Health Organization. WHO Expert Consultation on Rabies, third report (WHO Technical Report Series, No. 1012) [Internet]. Geneva: WHO; 2018. Available from: https:\/\/www.who.int\/publications\/i\/item\/WHO-TRS-101"},{"key":"2026091008070976178_j_almed-2025-0197_ref_003","doi-asserted-by":"crossref","unstructured":"Mani, RS, Dovih, DP, Ashwini, MA, Chattopadhyay, B, Harsha, PK, Garg, KM, et al.. Serological evidence of lyssavirus infection among Bats in Nagaland, a north-eastern state in India. Epidemiol Infect 2017;145:1635. https:\/\/doi.org\/10.1017\/S0950268817000310.","DOI":"10.1017\/S0950268817000310"},{"key":"2026091008070976178_j_almed-2025-0197_ref_004","doi-asserted-by":"crossref","unstructured":"Gunawardena, PS, Marston, DA, Ellis, RJ, Wise, EL, Karawita, AC, Breed, AC, et al.. Lyssavirus in Indian flying foxes, Sri Lanka. Emerg Infect Dis 2016;22:1456. https:\/\/doi.org\/10.3201\/EID2208.151986.","DOI":"10.3201\/eid2208.151986"},{"key":"2026091008070976178_j_almed-2025-0197_ref_005","unstructured":"World Organisation for Animal Health (WOAH). Manual of diagnostic tests and vaccines for terrestrial animals (Terrestrial manual). Twelfth: World Organisation for Animal Health (WOAH); 2023."},{"key":"2026091008070976178_j_almed-2025-0197_ref_006","unstructured":"An overview of antemortem and postmortem tests for diagnosis of human rabies. In: Rupprecht, CE, Fooks, AR, Abela-ridder, B, editors. Laboratory techniques in rabies, 5th ed. Geneva: World Health Organization; 2019, vol 1."},{"key":"2026091008070976178_j_almed-2025-0197_ref_007","doi-asserted-by":"crossref","unstructured":"Lodha, L, Ananda, AM, Mani, RS. Advancing rabies diagnosis: time for a new \u201cgold standard\u201d? Mol Diagn Ther 2025;29:153\u20139. https:\/\/doi.org\/10.1007\/s40291-024-00758-2.","DOI":"10.1007\/s40291-024-00758-2"},{"key":"2026091008070976178_j_almed-2025-0197_ref_008","unstructured":"Rupprecht, CE, Fooks, AR, Abela-Ridder, B. Laboratory techniques in rabies. Fifth: World Health Organization; 2018."},{"key":"2026091008070976178_j_almed-2025-0197_ref_009","doi-asserted-by":"crossref","unstructured":"Gigante, CM, Dettinger, L, Powell, JW, Seiders, M, Condori, REC, Griesser, R, et al.. Multi-site evaluation of the LN34\u00a0pan-lyssavirus real-time RT-PCR assay for post-mortem rabies diagnostics. PLoS One 2018;13. https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0197074.","DOI":"10.1371\/journal.pone.0197074"},{"key":"2026091008070976178_j_almed-2025-0197_ref_010","doi-asserted-by":"crossref","unstructured":"Wadhwa, A, Wilkins, K, Gao, J, Condori Condori, RE, Gigante, CM, Zhao, H, et al.. A pan-lyssavirus taqman real-time RT-PCR assay for the detection of highly variable rabies virus and other lyssaviruses. PLoS Neglected Trop Dis 2017;11:e0005258. https:\/\/doi.org\/10.1371\/JOURNAL.PNTD.0005258.","DOI":"10.1371\/journal.pntd.0005258"},{"key":"2026091008070976178_j_almed-2025-0197_ref_011","doi-asserted-by":"crossref","unstructured":"Mani, RS, Madhusudana, SN, Mahadevan, A, Reddy, V, Belludi, AY, Shankar, SK. Utility of real-time Taqman PCR for antemortem and postmortem diagnosis of human rabies. J\u00a0Med Virol 2014;86:1804\u201312. https:\/\/doi.org\/10.1002\/JMV.23814.","DOI":"10.1002\/jmv.23814"},{"key":"2026091008070976178_j_almed-2025-0197_ref_012","unstructured":"Dean, D, Abelseth, M, Atanasiu, P. The fluorescent antibody test. In: Meslin, F, Koprowsky, H, Kaplan, M, editors. WHO laboratory techniques in rabies, 4th ed. Geneva: WHO; 1996:88\u201395 pp."},{"key":"2026091008070976178_j_almed-2025-0197_ref_013","doi-asserted-by":"crossref","unstructured":"Nadin-Davis, SA, Sheen, M, Wandeler, AI. Development of real-time reverse transcriptase polymerase chain reaction methods for human rabies diagnosis. J\u00a0Med Virol 2009;81:1484\u201397. https:\/\/doi.org\/10.1002\/JMV.21547.","DOI":"10.1002\/jmv.21547"},{"key":"2026091008070976178_j_almed-2025-0197_ref_014","unstructured":"WHO (World Health Organization), Rupprecht, CE, Fooks, AR, Abela-Ridder, B. Laboratory techniques in rabies, 5th ed. World Health Organization; 2019, 2."},{"key":"2026091008070976178_j_almed-2025-0197_ref_015","doi-asserted-by":"crossref","unstructured":"Dettinger, L, Gigante, CM, Sellard, M, Seiders, M, Patel, P, Orciari, LA, et al.. Detection of apparent early rabies infection by LN34\u00a0pan-lyssavirus real-time RT-PCR assay in Pennsylvania. Viruses 2022;14:1845. https:\/\/doi.org\/10.3390\/v14091845.","DOI":"10.3390\/v14091845"},{"key":"2026091008070976178_j_almed-2025-0197_ref_016","doi-asserted-by":"crossref","unstructured":"Caraballo, DA, Lombardo, MA, Becker, P, Sabio, MS, Lema, C, Mart\u00ednez, LM, et al.. Evaluation of two real-time, TaqMan reverse transcription-PCR assays for detection of rabies virus in circulating variants from Argentina: influence of sequence variation. Viruses 2020;13:23. https:\/\/doi.org\/10.3390\/v13010023.","DOI":"10.3390\/v13010023"},{"key":"2026091008070976178_j_almed-2025-0197_ref_017","doi-asserted-by":"crossref","unstructured":"Minozzo, GA, Corona, TF, da Cruz, ECR, de Castro, WAC, Kmetiuk, LB, dos Santos, AP, et al.. Novel duplex RT\u2010qPCR for animal rabies surveillance. Transbound Emerg Dis 2022;69. https:\/\/doi.org\/10.1111\/tbed.14565.","DOI":"10.1111\/tbed.14565"},{"key":"2026091008070976178_j_almed-2025-0197_ref_018","doi-asserted-by":"crossref","unstructured":"Mauhay J, D, Saito, N, Kimitsuki, K, Mananggit M, R, Cruz J, L, Lagayan M, G, et al.. Molecular analysis of rabies virus using RNA extracted from used lateral flow devices. J\u00a0Clin Microbiol 2023;61. https:\/\/doi.org\/10.1128\/JCM.01543-22.","DOI":"10.1128\/jcm.01543-22"},{"key":"2026091008070976178_j_almed-2025-0197_ref_019","doi-asserted-by":"crossref","unstructured":"Demetria, C, Kimitsuki, K, Yahiro, T, Saito, N, Hashimoto, T, Khan, S, et al.. Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples. Trop Med Health 2023;51:17. https:\/\/doi.org\/10.1186\/s41182-023-00501-3.","DOI":"10.1186\/s41182-023-00501-3"},{"key":"2026091008070976178_j_almed-2025-0197_ref_020","doi-asserted-by":"crossref","unstructured":"Drzewniokov\u00e1, P, Marciano, S, Leopardi, S, Panzarin, V, De Benedictis, P. Comparison of pan-lyssavirus RT-PCRs and development of an improved protocol for surveillance of Non-RABV lyssaviruses. Viruses 2023;15:680. https:\/\/doi.org\/10.3390\/v15030680.","DOI":"10.3390\/v15030680"},{"key":"2026091008070976178_j_almed-2025-0197_ref_021","doi-asserted-by":"crossref","unstructured":"\u010cerne, D, Hostnik, P, Toplak, I, Presetnik, P, Maurer-Wernig, J, Kuhar, U. Discovery of a novel bat lyssavirus in a long-fingered bat (Myotis capaccinii) from Slovenia. PLoS Neglected Trop Dis 2023;17:e0011420. https:\/\/doi.org\/10.1371\/JOURNAL.PNTD.0011420.","DOI":"10.1371\/journal.pntd.0011420"},{"key":"2026091008070976178_j_almed-2025-0197_ref_022","doi-asserted-by":"crossref","unstructured":"Condori, RE, Niezgoda, M, Lopez, G, Matos, CA, Mateo, ED, Gigante, C, et al.. Using the LN34\u00a0pan-lyssavirus real-time RT-PCR assay for rabies diagnosis and rapid genetic typing from formalin-fixed human brain tissue. Viruses 2020;12. https:\/\/doi.org\/10.3390\/V12010120.","DOI":"10.3390\/v12010120"},{"key":"2026091008070976178_j_almed-2025-0197_ref_023","doi-asserted-by":"crossref","unstructured":"Blackburn, D, Minhaj, FS, Hammoud, RAl, Orciari, L, Miller, J, Maness, T, et al.. Human rabies\u00a0\u2013\u00a0texas, 2021. MMWR (Morb Mortal Wkly Rep) 2022;71:1547. https:\/\/doi.org\/10.15585\/MMWR.MM7149A2.","DOI":"10.15585\/mmwr.mm7149a2"},{"key":"2026091008070976178_j_almed-2025-0197_ref_024","doi-asserted-by":"crossref","unstructured":"Goravey, W, Husain, A, Ali, GA, Al Maslamani, MA, Ziglam, H. Antemortem diagnosis of human rabies: a case report. Clin Case Rep 2020;9:711\u20133. https:\/\/doi.org\/10.1002\/CCR3.3615.","DOI":"10.1002\/ccr3.3615"},{"key":"2026091008070976178_j_almed-2025-0197_ref_025","doi-asserted-by":"crossref","unstructured":"Peterson, D, Barbeau, B, McCaffrey, K, Gruninger, R, Eason, J, Burnett, C, et al.. Human rabies\u00a0\u2013\u00a0utah, 2018. MMWR Morb Mortal Wkly Rep 2020;69:121\u20134. https:\/\/doi.org\/10.15585\/MMWR.MM6905A1.","DOI":"10.15585\/mmwr.mm6905a1"},{"key":"2026091008070976178_j_almed-2025-0197_ref_026","doi-asserted-by":"crossref","unstructured":"Firpo, S, Piccirilli, MG, Urizar, R, Vitta, N, Hirmas Riade, SM, Leguizam\u00f3n, C, et al.. Human rabies by secondary transmission in Argentina, 2021. Diseases 2022;10:17. https:\/\/doi.org\/10.3390\/DISEASES10010017.","DOI":"10.3390\/diseases10010017"},{"key":"2026091008070976178_j_almed-2025-0197_ref_027","doi-asserted-by":"crossref","unstructured":"Wacharapluesadee, S, Tepsumethanon, V, Supavonwong, P, Kaewpom, T, Intarut, N, Hemachudha, T. Detection of rabies viral RNA by TaqMan real-time RT-PCR using non-neural specimens from dogs infected with rabies virus. J\u00a0Virol Methods 2012;184:109\u201312. https:\/\/doi.org\/10.1016\/J.JVIROMET.2012.05.013.","DOI":"10.1016\/j.jviromet.2012.05.013"}],"container-title":["Advances in Laboratory Medicine \/ Avances en Medicina de Laboratorio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.degruyterbrill.com\/document\/doi\/10.1515\/almed-2025-0197\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.degruyterbrill.com\/document\/doi\/10.1515\/almed-2025-0197\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:09:05Z","timestamp":1789027745000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.degruyterbrill.com\/document\/doi\/10.1515\/almed-2025-0197\/html"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,7]]},"references-count":27,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,7,7]]},"published-print":{"date-parts":[[2026,9,9]]}},"alternative-id":["10.1515\/almed-2025-0197"],"URL":"https:\/\/doi.org\/10.1515\/almed-2025-0197","relation":{},"ISSN":["2628-491X"],"issn-type":[{"value":"2628-491X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,7]]}}}