INVESTIGADORES
VOJNOV Adrian Alberto
artículos
Título:
Novel set of qPCR 16S rDNA primers for simultaneous detection of HLB 5 associated liberibacters
Autor/es:
INGRID GEORGINA ORCE; LORENA NOELIA SENDÍN; MARÍA ROSA MARANO; ADRIÁN ALBERTO VOJNOV; ATILIO PEDRO CASTAGNARO; MARÍA PAULA FILIPPONE
Revista:
SCIENTIA AGRICOLA
Editorial:
UNIV SAO PAOLO
Referencias:
Lugar: Piracicaba; Año: 2015 vol. 72 p. 252 - 259
ISSN:
0103-9016
Resumen:
Huanglongbing (HLB), a devastating citrus disease caused by the bacterium ?Candidatus Liberibacter spp.? is producing important economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacteria has severely hampered studies on the pathogen. 16S rDNA gene is a well characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species level. Quantitative Real Time PCR (qPCR) assays based on 16S rDNA genes are widely used for detection of ?Ca. Liberibacter spp.? in multiplex reactions. In this work, we have developed for first time one set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detect in a singleplex reaction all three bacterial species associated with HLB, and can differentiate Ca. Liberibacter asiaticus or africanus from americanus for their characteristic melting curves. The assay is extremely sensitive and were able to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value 3 times lower than tested methods for HLB diagnostic. The application of this fast, simple and efficient detection methodology could also be important in order to detect all species of HLB associated Liberibacters and could contribute to early pathogen detection, a crucial step to develop preventive strategies aimed to avoid the dissemination of this devastating disease in HLB-free areas.