دورية أكاديمية
أعرض في EDS

Rapid detection of West Nile and Dengue viruses from mosquito saliva by loop-mediated isothermal amplification and displaced probes.

التفاصيل البيبلوغرافية
العنوان: Rapid detection of West Nile and Dengue viruses from mosquito saliva by loop-mediated isothermal amplification and displaced probes.
المؤلفون: Kim D; Florida Medical Entomology Laboratory, University of Florida, Vero Beach, Florida, United States of America., DeBriere TJ; TrakitNow Inc., Columbia, South Carolina, United States of America., Eastmond BH; Florida Medical Entomology Laboratory, University of Florida, Vero Beach, Florida, United States of America., Alomar AA; Florida Medical Entomology Laboratory, University of Florida, Vero Beach, Florida, United States of America., Yaren O; Firebird Biomolecular Sciences LLC, Alachua, Florida, United States of America.; Foundation for Applied Molecular Evolution, Alachua, Florida, United States of America., McCarter J; Firebird Biomolecular Sciences LLC, Alachua, Florida, United States of America.; Foundation for Applied Molecular Evolution, Alachua, Florida, United States of America., Bradley KM; Foundation for Applied Molecular Evolution, Alachua, Florida, United States of America., Benner SA; Firebird Biomolecular Sciences LLC, Alachua, Florida, United States of America.; Foundation for Applied Molecular Evolution, Alachua, Florida, United States of America., Alto BW; Florida Medical Entomology Laboratory, University of Florida, Vero Beach, Florida, United States of America., Burkett-Cadena ND; Florida Medical Entomology Laboratory, University of Florida, Vero Beach, Florida, United States of America.
المصدر: PloS one [PLoS One] 2024 Feb 23; Vol. 19 (2), pp. e0298805. Date of Electronic Publication: 2024 Feb 23 (Print Publication: 2024).
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
أسماء مطبوعة: Original Publication: San Francisco, CA : Public Library of Science
مواضيع طبية MeSH: Dengue Virus*/genetics , Culicidae* , Arboviruses* , Nucleic Acid Amplification Techniques* , Molecular Diagnostic Techniques*, Animals ; Humans ; Saliva ; Mosquito Vectors ; RNA ; Sucrose
مستخلص: Arthropod-borne viruses are major causes of human and animal disease, especially in endemic low- and middle-income countries. Mosquito-borne pathogen surveillance is essential for risk assessment and vector control responses. Sentinel chicken serosurveillance (antibody testing) and mosquito pool screening (by RT-qPCR or virus isolation) are currently used to monitor arbovirus transmission, however substantial time lags of seroconversion and/or laborious mosquito identification and RNA extraction steps sacrifice their early warning value. As a consequence, timely vector control responses are compromised. Here, we report on development of a rapid arbovirus detection system whereby adding sucrose to reagents of loop-mediated isothermal amplification with displaced probes (DP-LAMP) elicits infectious mosquitoes to feed directly upon the reagent mix and expectorate viruses into the reagents during feeding. We demonstrate that RNA from pathogenic arboviruses (West Nile and Dengue viruses) transmitted in the infectious mosquito saliva was detectable rapidly (within 45 minutes) without RNA extraction. Sucrose stabilized viral RNA at field temperatures for at least 48 hours, important for transition of this system to practical use. After thermal treatment, the DP-LAMP could be reliably visualized by a simple optical image sensor to distinguish between positive and negative samples based on fluorescence intensity. Field application of this technology could fundamentally change conventional arbovirus surveillance methods by eliminating laborious RNA extraction steps, permitting arbovirus monitoring from additional sites, and substantially reducing time needed to detect circulating pathogens.
Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: O.Y., S.A.B., and their institutions own intellectual property associated with DP-LAMP assay.
(Copyright: © 2024 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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معلومات مُعتمدة: R44 CK000578 United States CK NCEZID CDC HHS
المشرفين على المادة: 63231-63-0 (RNA)
57-50-1 (Sucrose)
SCR Protocol: LAMP assay
تواريخ الأحداث: Date Created: 20240223 Date Completed: 20240226 Latest Revision: 20240310
رمز التحديث: 20240310
مُعرف محوري في PubMed: PMC10889885
DOI: 10.1371/journal.pone.0298805
PMID: 38394282
قاعدة البيانات: MEDLINE
الوصف
تدمد:1932-6203
DOI:10.1371/journal.pone.0298805