1.Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification.
Guan LI ; Kai NIE ; Dan ZHANG ; Xinna LI ; Yanqun WANG ; Wenjie TAN ; Xuejun MA
Chinese Journal of Virology 2015;31(3):269-275
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.
Coronavirus Infections
;
virology
;
DNA Primers
;
genetics
;
Humans
;
Middle East Respiratory Syndrome Coronavirus
;
classification
;
genetics
;
isolation & purification
;
Nucleic Acid Amplification Techniques
;
methods
;
Reverse Transcription
2.SARS-like virus in the Middle East: a truly bat-related coronavirus causing human diseases.
Protein & Cell 2012;3(11):803-805
Animals
;
Chiroptera
;
virology
;
Coronavirus
;
classification
;
genetics
;
isolation & purification
;
Coronavirus Infections
;
transmission
;
virology
;
Databases, Genetic
;
Genome, Viral
;
Humans
;
Male
;
Middle Aged
;
Middle East
;
Phylogeny
;
SARS Virus
;
classification
;
genetics
;
isolation & purification
;
Severe Acute Respiratory Syndrome
;
virology
3.Isolation of Middle East Respiratory Syndrome Coronavirus from a Patient of the 2015 Korean Outbreak.
Wan Beom PARK ; Nak Jung KWON ; Pyoeng Gyun CHOE ; Su Jin CHOI ; Hong Sang OH ; Sang Min LEE ; Hyonyong CHONG ; Jong Il KIM ; Kyoung Ho SONG ; Ji Hwan BANG ; Eu Suk KIM ; Hong Bin KIM ; Sang Won PARK ; Nam Joong KIM ; Myoung Don OH
Journal of Korean Medical Science 2016;31(2):315-320
During the 2015 outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) in Korea, 186 persons were infected, resulting in 38 fatalities. We isolated MERS-CoV from the oropharyngeal sample obtained from a patient of the outbreak. Cytopathic effects showing detachment and rounding of cells were observed in Vero cell cultures 3 days after inoculation of the sample. Spherical virus particles were observed by transmission electron microscopy. Full-length genome sequence of the virus isolate was obtained and phylogenetic analyses showed that it clustered with clade B of MERS-CoV.
Animals
;
Cercopithecus aethiops
;
Coronavirus Infections/*diagnosis/epidemiology/*virology
;
Disease Outbreaks
;
Humans
;
Microscopy, Electron
;
Middle East Respiratory Syndrome Coronavirus/classification/genetics/*isolation & purification/ultrastructure
;
Phylogeny
;
Polymerase Chain Reaction
;
RNA, Viral/analysis/chemistry/metabolism
;
Republic of Korea/epidemiology
;
Sequence Analysis, RNA
;
Vero Cells