INVESTIGADORES
GARCIA Maria Laura
artículos
Título:
A highly sensitive heminested RT-PCR assay for the detection of citrus psorosis virus targeted to a conserved region of the genome
Autor/es:
G.G. LEGARRETA, M.L. GARCÍA, N. COSTA ,O. GRAU
Revista:
JOURNAL OF VIROLOGICAL METHODS
Editorial:
ELSEVIER SCIENCE BV
Referencias:
Lugar: Amsterdam; Año: 2000 vol. 84 p. 15 - 22
ISSN:
0166-0934
Resumen:
Psorosis is a widespread and damaging disease of citrus in many parts of the world. The causal agent is a multipartite virus with RNA genome present in very low concentration in infected citrus tissue. Diagnosis is made by biological indexing on indicator citrus seedlings, but it is a slow and costly procedure and therefore it is not used generally. No sensitive wide-spectrum assay for Citrus Psorosis virus (CPsV) has been reported based on RT-PCR. A highly sensitive heminested RT-PCR assay is described for the detection of CPsV. Fragments of 313 bp amplified from RNA 1 of different isolates were cloned and sequenced. Very high homology was found among six isolates from the citrus producing region of Argentina: 96.6–100% in nucleotide sequence. The consensus sequence obtained was used for the design of the primers for heminested PCR assay. It has been tested on different Argentine isolates, employing various methods for RNA extraction from infected tissue. This test is able to detect CPsV in dilutions of 1010 of the original sample. © 2000 Elsevier Science B.V. All rights reserved