Visual Detection of Human Coronavirus NL63 by Reverse Transcription Loop-Mediated Isothermal Amplification.
- Author:
Heyuan GENG
;
Shengqiang WANG
;
Xiaoqian XIE
;
Yu XIAO
;
Ting ZHANG
;
Wenjie TAN
;
Chuan SU
- Publication Type:Journal Article
- MeSH:
Colorimetry;
methods;
Coronavirus Infections;
diagnosis;
virology;
Coronavirus NL63, Human;
genetics;
isolation & purification;
DNA Primers;
genetics;
Humans;
Nucleic Acid Amplification Techniques;
methods;
Reverse Transcription;
Sensitivity and Specificity
- From:
Chinese Journal of Virology
2016;32(1):56-61
- CountryChina
- Language:Chinese
-
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.
