Identification and detection of Cordyceps sinensis with LAMP / 中草药

ABSTRACT

Objective: The method of loop-mediated isothermal amplification (LAMP) was employed to detect and identify Cordyceps sinensis rapidly. Methods: Specific LAMP primers were designed according to the CS2 serine protease (csp2) gene of Cordyceps sinensis. C. sinensis DNA was extracted using CTAB method. The reaction conditions of LAMP were optimized. Specificity of LAMP reaction was validated by six different strains and using restriction enzyme Taq1 digested the LAMP products. Sensitivity of LAMP was tested with diluted C. sinensis solution with 10-fold gradient. LAMP products were shown by gel electrophoresis or adding SYBR Green I. Results: The method of LAMP for detecting C. sinensis was effective and specific. The detection limit of LAMP assay was up to 6 pg/mL. Conclusion: LAMP protocol is a promising method for the identification and detection of C. sinensis and Chinese materia medica as well.