{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T16:56:14Z","timestamp":1787504174898,"version":"build-2736575974"},"reference-count":18,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2024,3,13]],"date-time":"2024-03-13T00:00:00Z","timestamp":1710288000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,3,13]],"date-time":"2024-03-13T00:00:00Z","timestamp":1710288000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000180","name":"U.S. Department of Homeland Security","doi-asserted-by":"crossref","award":["HSHQDC 16-A-B0006"],"award-info":[{"award-number":["HSHQDC 16-A-B0006"]}],"id":[{"id":"10.13039\/100000180","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["P20GM130448"],"award-info":[{"award-number":["P20GM130448"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2024,3,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Rift Valley Fever phlebovirus (RVFV) is a mosquito-borne zoonotic pathogen that causes major agricultural and public health problems in Africa and the Arabian Peninsula. It is considered a potential agro-bioterrorism agent for which limited countermeasures are available. To address diagnostic needs, here we describe a rapid and sensitive molecular method immediately employable at sites of suspected outbreaks in animals that commonly precede outbreaks in humans. The strategy involves the concurrent detection of two of the three RVFV genome segments (large and medium) using reverse transcription insulated isothermal PCR (RT-iiPCR) performed on a portable, touch screen nucleic acid analyzer, POCKIT. The analytical sensitivity for both the RT-iiPCR and a laboratory-based L and M multiplex reverse transcription real-time PCR assay was estimated at approximately 0.1\u20133 copies\/reaction using synthetic RNA or viral RNA. The diagnostic sensitivity and specificity of detection of RVFV on the POCKIT, determined using sera from sheep and cattle (\n                      <jats:italic>n<\/jats:italic>\n                      = 181) experimentally infected with two strains of RVFV (SA01 and Ken06), were 93.8% and 100% (kappa = 0.93), respectively. Testing of ruminant field sera (\n                      <jats:italic>n<\/jats:italic>\n                      = 193) in two locations in Africa demonstrated 100% diagnostic sensitivity and specificity. We conclude that the POCKIT dual-gene RVFV detection strategy can provide reliable, sensitive, and specific point-of-need viral RNA detection. Moreover, the field detection of RVFV in vectors or susceptible animal species can aid in the surveillance and epidemiological studies to better understand and control RVFV outbreaks.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The content of this manuscript is of interest to the diverse readership of the\n                      <jats:italic>Journal of Clinical Microbiology<\/jats:italic>\n                      , including research scientists, diagnosticians, healthcare professionals, and policymakers. Rift Valley Fever virus (RVFV) is a zoonotic mosquito-borne pathogen that causes major agricultural and public health problems. Current and most sensitive diagnostic approaches that are molecular-based are performed in highly specialized molecular diagnostic laboratories. To address diagnostic needs, we developed a novel, rapid, and sensitive molecular method using a portable PCR machine, POCKIT, capable of immediate deployment at sites of suspected outbreaks. Here, we demonstrate that field-deployable RVFV detection can provide reliable, sensitive, and specific point-of-need viral RNA detection that could be used for diagnostic investigations and epidemiological studies, and can be performed in the field.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.00430-23","type":"journal-article","created":{"date-parts":[[2024,2,2]],"date-time":"2024-02-02T09:00:35Z","timestamp":1706864435000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["Rift Valley Fever virus M and L genome segment detection: a comparison of field-deployable reverse transcription insulated isothermal PCR (RT-iiPCR) and laboratory-based multiplex reverse transcription real-time PCR"],"prefix":"10.1128","volume":"62","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5644-022X","authenticated-orcid":true,"given":"Jessie D.","family":"Trujillo","sequence":"first","affiliation":[{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, Diagnostic Medicine\/Pathobiology, Kansas State University, Manhattan, Kansas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William C.","family":"Wilson","sequence":"additional","affiliation":[{"name":"Foreign Arthropod-Borne Animal Diseases Research Unit (FABADRU), USDA Agricultural Research Service (ARS), Manhattan, Kansas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony","family":"Craig","sequence":"additional","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Vectors and Vector-Borne Diseases Research Programme, Pretoria, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carien","family":"Van den Bergh","sequence":"additional","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Vectors and Vector-Borne Diseases Research Programme, Pretoria, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Wang","sequence":"additional","affiliation":[{"name":"Research and development, GeneReach USA, Lexington, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Thompson","sequence":"additional","affiliation":[{"name":"Department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Swanepoel","sequence":"additional","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Vectors and Vector-Borne Diseases Research Programme, Pretoria, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Igor","family":"Morozov","sequence":"additional","affiliation":[{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, Diagnostic Medicine\/Pathobiology, Kansas State University, Manhattan, Kansas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7308-5672","authenticated-orcid":true,"given":"Juergen A.","family":"Richt","sequence":"additional","affiliation":[{"name":"Center of Excellence for Emerging and Zoonotic Animal Diseases, Diagnostic Medicine\/Pathobiology, Kansas State University, Manhattan, Kansas, USA"},{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Vectors and Vector-Borne Diseases Research Programme, Pretoria, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-017-3358-5"},{"key":"e_1_3_4_3_2","first-page":"1037","article-title":"Rift Valley fever","volume":"2","author":"Swanepoel R","year":"2004","unstructured":"Swanepoel R, Coetzer J. 2004. Rift Valley fever. Infectious diseases of livestock 2:1037\u20131070.","journal-title":"Infectious diseases of livestock"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1159\/000373924"},{"key":"e_1_3_4_5_2","article-title":"Vaccines and diagnostics for transboundary animal diseases international symposium, Ames, Iowa, September 2012: proceedings","volume":"135","author":"Roth JA","year":"2013","unstructured":"Roth JA, Richt JA, Morozov IA. 2013. Vaccines and diagnostics for transboundary animal diseases international symposium, Ames, Iowa, September 2012: proceedings. 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