%0 Journal Article
%T Rapid and sensitive detection of Citrus Bacterial Canker by loop-mediated isothermal amplification combined with simple visual evaluation methods
%A Luciano A Rigano
%A Mar¨ªa R Marano
%A Atilio P Castagnaro
%A Alexandre Do Amaral
%A Adrian A Vojnov
%J BMC Microbiology
%D 2010
%I BioMed Central
%R 10.1186/1471-2180-10-176
%X A loop-mediated isothermal amplification assay for the diagnosis of Citrus Bacterial Canker (CBC-LAMP) was developed and evaluated. DNA samples were obtained from infected plants or cultured bacteria. A typical ladder-like pattern on gel electrophoresis was observed in all positive samples in contrast to the negative controls. In addition, amplification products were detected by visual inspection using SYBRGreen and using a lateral flow dipstick, eliminating the need for gel electrophoresis. The sensitivity and specificity of the assay were evaluated in different conditions and using several sample sources which included purified DNA, bacterium culture and infected plant tissue. The sensitivity of the CBC-LAMP was 10 fg of pure Xcc DNA, 5 CFU in culture samples and 18 CFU in samples of infected plant tissue. No cross reaction was observed with DNA of other phytopathogenic bacteria. The assay was capable of detecting CBC-causing strains from several geographical origins and pathotypes.The CBC-LAMP technique is a simple, fast, sensitive and specific method for the diagnosis of Citrus Bacterial Canker. This method can be useful in the phytosanitary programs of the citrus industry worldwide.Citrus Bacterial Canker is an economic important disease in several countries, and causes great losses in fruit production and its subsidiaries [1]. There are three types of Citrus Bacterial Canker identified that have different genotypes and posses variations in host range among citrus plants. The type A CBC originating from Asia, is caused by Xanthomonas citri subsp. citri, this is the most destructive and widespread variant of the disease with a host range that includes all citrus cultivars [2]. The CBC types B and C are caused by Xanthomonas fuscans subsp. aurantifolii strains B and C, respectively. Those bacteria are limited in host range and are geographically restricted to South America. Type B CBC is present only in Argentina, Uruguay and Paraguay and is found primarily on le
%U http://www.biomedcentral.com/1471-2180/10/176