title: Lyophilized visually readable loop-mediated isothermal reverse transcriptase nucleic acid amplification test for detection Ebola Zaire RNA.
abstract: Recent viral outbreaks highlight the need for reliable, yet broadly deployable diagnostics for detection of epidemic and emerging pathogens. In this study we designed and optimized methods to visually detect viral nucleic acid by isothermal amplification and SYBR dye intercalation. We designed and tested loop-mediated isothermal amplification (LAMP) primers and lyophilized reactions to optimize the detection of Zaire Ebola Virus (ZEBOV) and further evolved the LAMP platform to allow room-temperature storage for deployment in resource limited settings. Our results demonstrated excellent sensitivity and specificity for viral nucleic acid sequences with lower limits of detection of less than 100 copies. Moreover, lyophilized reaction mixtures retained activity for prolonged periods under dry conditions at room temperature. This approach offers a way for detection of emerging viruses in resource limited settings.
authors.affiliation: University of California, Department of Medicine, Division of Infectious Disease, 9500 Gilman Drive # 0711, La Jolla, San Diego, CA 92093-0711, USA. Electronic address: christoph.c.carter@gmail.com.
authors.forename: Christoph
authors.initials: C
authors.lastname: Carter
authors.affiliation: University of California, Electrical and Computer Engineering, 9500 Gilman Drive, Jacobs Hall, Room 4805, La Jolla, San Diego, CA, USA. Electronic address: kakrami@ucsd.edu.
authors.forename: Kevan
authors.initials: K
authors.lastname: Akrami
authors.affiliation: University of California, Department of Medicine, Division of Infectious Disease, 9500 Gilman Drive # 0711, La Jolla, San Diego, CA 92093-0711, USA; University of California, Electrical and Computer Engineering, 9500 Gilman Drive, Jacobs Hall, Room 4805, La Jolla, San Diego, CA, USA. Electronic address: drewhall@ucsd.edu.
authors.forename: Drew
authors.initials: D
authors.lastname: Hall
authors.affiliation: University of California, Electrical and Computer Engineering, 9500 Gilman Drive, Jacobs Hall, Room 4805, La Jolla, San Diego, CA, USA. Electronic address: davey@ucsd.edu.
authors.forename: Davey
authors.initials: D
authors.lastname: Smith
authors.affiliation: University of California, Electrical and Computer Engineering, 9500 Gilman Drive, Jacobs Hall, Room 4805, La Jolla, San Diego, CA, USA. Electronic address: earonoffspencer@ucsd.edu.
authors.forename: Eliah
authors.initials: E
authors.lastname: Aronoff-Spencer
mesh_terms: D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D005612:Freeze Drying; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic
keywords: Ebola; Lyophilized; Nucleic-acid-amplification; Reverse-transcriptase