Rapid detection of variola virus by revolution transcription loop mediated isothermal amplification method / 中华实验和临床病毒学杂志

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.