title: Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus.
abstract: During dengue outbreaks, acute diagnosis at the patient's point of need followed by appropriate supportive therapy reduces morbidity and mortality. To facilitate needed diagnosis, we developed and optimized a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay that detects all 4 serotypes of dengue virus (DENV). We used a quencher to reduce nonspecific amplification. The assay does not require expensive thermocyclers, utilizing a simple water bath to maintain the reaction at 63 °C. Results can be visualized using UV fluorescence, handheld readers, or lateral flow immunochromatographic tests. We report a sensitivity of 86.3% (95% confidence interval [CI], 72.7-94.8%) and specificity of 93.0% (95% CI, 83.0-98.1%) using a panel of clinical specimens characterized by DENV quantitative reverse transcription-polymerase chain reaction. This pan-serotype DENV RT-LAMP can be adapted to field-expedient formats where it can provide actionable diagnosis near the patient's point of need.
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Allison L
authors.initials: AL
authors.lastname: Dauner
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Indrani
authors.initials: I
authors.lastname: Mitra
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Theron
authors.initials: T
authors.lastname: Gilliland
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Sajeewane
authors.initials: S
authors.lastname: Seales
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA. Electronic address: subhamoy.pal.ctr@mail.mil.
authors.forename: Subhamoy
authors.initials: S
authors.lastname: Pal
authors.affiliation: National Chung Hsing University, Taichung, Taiwan.
authors.forename: Shih-Chun
authors.initials: SC
authors.lastname: Yang
authors.affiliation: US Naval Medical Research Unit-6, Iquitos, Peru.
authors.forename: Carolina
authors.initials: C
authors.lastname: Guevara
authors.affiliation: National Chung Hsing University, Taichung, Taiwan.
authors.forename: Jiann-Hwa
authors.initials: JH
authors.lastname: Chen
authors.affiliation: National Chung Hsing University, Taichung, Taiwan.
authors.forename: Yung-Chuan
authors.initials: YC
authors.lastname: Liu
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Tadeusz J
authors.initials: TJ
authors.lastname: Kochel
authors.affiliation: Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
authors.forename: Shuenn-Jue L
authors.initials: SJ
authors.lastname: Wu
mesh_terms: D003715:Dengue / Q000175:diagnosis* / Q000821:virology*; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000378:metabolism; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D065288:Serogroup; D013696:Temperature
chemical_list: D012367:RNA, Viral
keywords: Dengue; Loop-mediated isothermal amplification; RT-LAMP