Visual Detection of Human Coronavirus NL63 by Reverse Transcription Loop-Mediated Isothermal Amplification / 病毒学报
Chinese Journal of Virology
;
(6): 56-61, 2016.
Article
in Chinese
| WPRIM
| ID: wpr-296217
ABSTRACT
A simple and sensitive assay for rapid detection of human coronavirus NL63 (HCoV-NL63) was developed by colorimetic reverse transcription loop-mediated isothermal amplification (RT-LAMP). The method employed six specially designed primers that recognized eight distinct regions of the HCoV-NL63 nucleocapsid protein gene for amplification of target sequences under isothermal conditions at 63 degrees C for 1 h Amplification of RT-LAMP was monitored by addition of calcein before amplification. A positive reaction was confirmed by change from light-brown to yellow-green under visual detection. Specificity of the RT-LAMP assay was validated by cross-reaction with different human coronaviruses, norovirus, influenza A virus, and influenza B virus. Sensitivity was evaluated by serial dilution of HCoV-NL63 RNA from 1.6 x 10(9) to 1.6 x 10(1) per reaction. The RT-LAMP assay could achieve 1,600 RNA copies per reaction with high specificity. Hence, our colorimetric RT-LAMP assay could be used for rapid detection of human coronavirus NL63.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Virology
/
Sensitivity and Specificity
/
DNA Primers
/
Coronavirus Infections
/
Colorimetry
/
Nucleic Acid Amplification Techniques
/
Reverse Transcription
/
Diagnosis
/
Coronavirus NL63, Human
/
Genetics
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Virology
Year:
2016
Type:
Article
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