Rapid detection of variola virus by revolution transcription loop mediated isothermal amplification method
10.3760/cma.j.issn.1003-9279.2019.03.018
- VernacularTitle: 基于LAMP法快速检测天花病毒
- Author:
Longfei FENG
1
;
Bing DU
2
;
Yingjie HUANG
1
;
Yang CAO
3
;
Luping CHEN
3
;
Xinhong LIANG
1
;
Huan LI
3
;
Jing YUAN
3
Author Information
1. Henan Institute of Science and Technology, School of Food Science, Xinxiang, Henan 453003, China
2. Beijing City University, Beijing 100071, China
3. Institute of Disease Control and Prevention, China People′s Liberation Army, Beijing 100071, China
- Publication Type:Journal Article
- Keywords:
Variola virus;
Loop-mediated isothermal amplification;
Rapid detection
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(3):314-318
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish the loop-mediated isothermal amplification (LAMP) method for detection of variola virus.
Methods:One set of primers were designed for recognizing 5 distinct sequences on variola virus-specific gene HA. To optimize the reaction temperature and primers screening, and the sensitivity and specificity of this method for variola virus (VARV) detection were evaluated.
Results:Rapid detection of variola virus by LAMP assay was completed within 60 min at 63 ℃. The sensitivity of LAMP with detection limits of 1 pg/μl was 10 times higher than that of PCR, that is, the LAMP sensitivity was 3.37×105 copies/μl, and the PCR sensitivity was 3.37×106 copies/μl. and the result of 2 kinds of other virus were negative, showing that it had a good specificity.
Conclusions:The method reported here demonstrates a potential and valuable means for detection of VARV. The LAMP assay is suitable for wide-area sample screening and on-site detection of variola virus in grassroots units, for on-site and primary quarantine, medical units for rapid diagnosis.
