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Development of Loop-Mediated Isothermal Amplification for Detection of subsp. in Sugarcane

Jing Liu, Liping Xu, Jinlong Guo, Rukai Chen, Michael Paul Grisham, Youxiong Que
BioMed Research International 2013 v.2013 pp. 1-8
DNA, Leifsonia xyli subsp. xyli, Saccharum, bacteria, color, internodes, loop-mediated isothermal amplification, microbial detection, pathogen identification, polymerase chain reaction, ratoon stunting disease, sap, sugarcane
Ratoon stunt, caused by the xylem-limited coryneform bacterium Leifsonia xyli subsp. xyli (Lxx), is prevalent in most sugarcane-planting countries. Because the disease does not cause characteristic external symptoms, a laboratory-based technique is needed for accurate diagnosis. Based on loop-mediated isothermal amplification (LAMP), we developed a method for detecting Lxx. The major advantages of the LAMP method are visual judgment by colour, reduced time (60 min), and not needing a costly PCR apparatus and gel scanner. In the present study, positive and negative samples detected by the LAMP method were clearly distinguishable. When total DNA extracted from internode juice was used as the template, the sensitivity of LAMP was 10 times higher than that of the conventional PCR detection. The LAMP assay is a highly specific, rapid and sensitive method for the diagnosis of ratoon stunt caused by Lxx in sugarcane.