REDBIO México 2010

Folio: 263
Tipo de presentación: Oral
Tema: Estrés Biótico
Este trabajo se presentará en "Presentaciones Libres 5: Estrés" el Martes dentro del horario 14:20 a 16:20

Rapid and Sensitive Xanthomonas citri detection by LAMP combined with naked eye evaluation methods

Luciano A. Rigano * (1), Adrian A. Vojnov (1).
* Autor para correspondencia (Corresponding author) avojnov@fundacioncassara.org.ar

1- FPC-Centro Milstein, Saladillo 2468, C1440FFX, Buenos Aires, Argentina

RESUMEN (ABSTRACT)
"Xanthomomas citri pv. citri is the bacterial pathogen that causes the canker disease in citrus plants worldwide. In southamérica this disease is an important problem for citrus industry in Argentina and Brazil. Detection methods based on antibodies or PCR has been developed previously, but this approaches are time consuming, laborious and requires expensive laboratory equipment. Loop mediated isothermal amplification (LAMP), a novel isothermal DNA amplification technique, is very sensitive, specific, fast and does not require specialized laboratory equipment. A loop mediated isothermal amplification assay for the detection of Xanthomonas citri pv. citri (Xac-LAMP) was developed and evaluated. The assay was optimized for the amplification of a portion of the pthA gene, a patogenicity determinant of Xac. A tipical ladder like pattern on gel electrophoresis was observed in all positive samples. Also amplification products were detected by visual inspection using SybrGreen and using a lateral flow dipstick, eliminating the need of time consuming gel electrophoresis. The sensibility and specificity of the assay were evaluated in different conditions. Not cross reaction was observed with pure DNA of other phytopathogenic bacteria. The Xac-LAMP assay is a simple, fast, sensitive and specific alternative for the accurate detection of Xanthomonas citri pv. citri."