Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus

被引：18

作者：

Dauner, Allison L. ^{[1
]}

Mitra, Indrani ^{[1
]}

Gilliland, Theron, Jr. ^{[1
]}

Seales, Sajeewane ^{[1
]}

Pal, Subhamoy ^{[1
]}

Yang, Shih-Chun ^{[2
]}

Guevara, Carolina ^{[3
]}

Chen, Jiann-Hwa ^{[2
]}

Liu, Yung-Chuan ^{[2
]}

Kochel, Tadeusz J. ^{[1
]}

Wu, Shuenn-Jue L. ^{[1
]}

机构：

[1] Naval Med Res Ctr, Silver Spring, MD 20910 USA

[2] Natl Chung Hsing Univ, Taichung 40227, Taiwan

[3] US Naval Med Res Unit 6, Iquitos, Peru

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2015年 / 83卷 / 01期

关键词：

Dengue; Loop-mediated isothermal amplification; RI-LAMP; JAPANESE ENCEPHALITIS-VIRUS; WEST-NILE-VIRUS; RAPID DETECTION; SENSITIVE DETECTION; NUCLEIC-ACID; VIRAL-RNA; RT-LAMP; DIAGNOSIS; DNA; DIFFERENTIATION;

D O I：

10.1016/j.diagmicrobio.2015.05.004

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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 (RI-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 degrees 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% Cl. 83.0-98.1%) using a panel of clinical specimens characterized by DENV quantitative reverse transcription-polymerase chain reaction. This pan-serotype DENV RI-LAMP can be adapted to field-expedient formats where it can provide actionable diagnosis near the patient's point of need. (C) 2015 Elsevier Inc. All rights reserved.

引用

页码：30 / 36

页数：7

共 45 条

[41] Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus [J].