title: [Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification].
abstract: A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.
authors.forename: Guan
authors.initials: G
authors.lastname: Li
authors.forename: Kai
authors.initials: K
authors.lastname: Nie
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.forename: Xinna
authors.initials: X
authors.lastname: Li
authors.forename: Yanqun
authors.initials: Y
authors.lastname: Wang
authors.forename: Wenjie
authors.initials: W
authors.lastname: Tan
authors.forename: Xuejun
authors.initials: X
authors.lastname: Ma
mesh_terms: D018352:Coronavirus Infections / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D065207:Middle East Respiratory Syndrome Coronavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D048348:Reverse Transcription
chemical_list: D017931:DNA Primers