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Development of a Loop-Mediated Isothermal Amplification Procedure as a Sensitive and Rapid Method for Detection of ‘Candidatus Liberibacter solanacearum’ in Potatoes and Psyllids

September 2012 , Volume 102 , Number  9
Pages  899 - 907

Aravind Ravindran, Julien Levy, Elizabeth Pierson, and Dennis C. Gross

First and fourth authors: Department of Plant Pathology and Microbiology, and second and third authors: Department of Horticultural Sciences, Texas A&M University, College Station 77843.

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Accepted for publication 12 May 2012.

This study reports the development of a loop-mediated isothermal amplification procedure (LAMP) for polymerase chain reaction (PCR)-based detection of ‘Candidatus Liberibacter solanacearum’, the bacterial causal agent of potato zebra chip (ZC) disease. The 16S rDNA gene of ‘Ca. Liberibacter solanacearum’ was used to design a set of six primers for LAMP PCR detection of the bacterial pathogen in potato plants and the psyllid vector. The advantage of the LAMP method is that it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Positive LAMP results can be visualized directly as a precipitate. The LAMP strategy reported here reliably detected ‘Ca. Liberibacter solanacearum’ and the closely related species ‘Ca. Liberibacter asiaticus’, the causative agent of huanglongbing disease of citrus, in plant DNA extracts. Although not as sensitive as quantitative real-time PCR, LAMP detection was equivalent to conventional PCR in tests of ZC-infected potato plants from the field. Thus, the LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting ‘Ca. Liberibacter’ pathogens in psyllids and field-grown potato plants and tubers.

© 2012 The American Phytopathological Society