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. Key words: Variola virus; Loop-mediated isothermal amplification; Rapid detection
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