Development and Evaluation of Isothermal Amplification Methods for Rapid Detection of Lethal Species.

In the present work, loop-mediated isothermal amplification (LAMP) and hyperbranched rolling circle amplification (HRCA) methods were developed to detect and distinguish different lethal species. Specific LAMP primers and HRCA padlock probes for species-specific identification and a set of universal LAMP primers for lethal species were designed and tested. The results indicated that the LAMP-based assay was able to discriminate introclade lethal species but was not able to discriminate intraclade species perfectly, while the HRCA-based assay could discriminate whether introclade or intraclade species. The universal LAMP primers were positive for 10 lethal species of section and negative for 16 species of outside section . The detection limits of LMAP and HRCA were 10 and 1 pg of genomic DNA per reaction, respectively. In conclusion, the two methods could be rapid, specific, sensitive and low-cost tools for the identification of lethal species.