title: Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly From Blood.
abstract: A molecular diagnostic method for robust detection of Ebola virus (EBOV) at the point of care (POC) directly from blood samples is described. This assay is based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) of the glycoprotein gene of EBOV. Complete reaction formulations were lyophilized in 0.2-mL polymerase chain reaction tubes. RT-LAMP reactions were performed on a battery-operated isothermal instrument. Limit of detection of this RT-LAMP assay was 2.8 × 102plaque-forming units (PFU)/test and 1 × 103PFU/test within 40 minutes for EBOV-Kikwit and EBOV-Makona, respectively. This assay was found to be specific for the detection of EBOV, as no nonspecific amplification was detected in blood samples spiked with closely related viruses and other pathogens. These results showed that this diagnostic test can be used at the point of care for rapid and specific detection of EBOV directly from blood with high sensitivity within 40 minutes.
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Benzine
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Brown
authors.affiliation: Department of Microbiology and Immunology Galveston National Laboratory, University of Texas Medical Branch at Galveston.
authors.forename: Krystle N
authors.initials: KN
authors.lastname: Agans
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Ronald
authors.initials: R
authors.lastname: Godiska
authors.affiliation: Department of Microbiology and Immunology Galveston National Laboratory, University of Texas Medical Branch at Galveston.
authors.forename: Chad E
authors.initials: CE
authors.lastname: Mire
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Krishne
authors.initials: K
authors.lastname: Gowda
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Brandon
authors.initials: B
authors.lastname: Converse
authors.affiliation: Department of Microbiology and Immunology Galveston National Laboratory, University of Texas Medical Branch at Galveston.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Geisbert
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: David A
authors.initials: DA
authors.lastname: Mead
authors.affiliation: Lucigen Corp., Middleton, Wisconsin.
authors.forename: Yogesh
authors.initials: Y
authors.lastname: Chander
mesh_terms: D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D025202:Molecular Diagnostic Techniques; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral
keywords: Ebola virus; RT-LAMP; diagnostic test; isothermal amplification; point of care