Skip navigation

Development of an asymmetric PCR-ELISA typing method for citrus tristeza virus based on the coat protein gene

Development of an asymmetric PCR-ELISA typing method for citrus tristeza virus based on the coat protein gene

Nolasco, G., Santos, C., Silva, G. ORCID: 0000-0001-5544-2947 and Fonseca, F. (2008) Development of an asymmetric PCR-ELISA typing method for citrus tristeza virus based on the coat protein gene. Journal of Virological Methods, 155 (2). pp. 97-108. ISSN 0166-0934 (doi:10.1016/j.jviromet.2008.09.030)

Full text not available from this repository.

Abstract

The coat protein gene of isolates of citrus tristeza virus (CTV) from 20 citrus-producing regions around the world was amplified by RT-PCR, TA cloned, and characterized by SSCP. Haplotypes that produced different patterns within each geographic region were sequenced and a database of 153 accessions of CTV was assembled. Phylogenetic analysis revealed the existence of seven well-defined clusters (Coefficient of differentiation 0.78). An asymmetric PCR-ELISA typing (APET) assay was developed in the frame of this clustering pattern using a set of eight hybridisation probes. The membership of any unknown haplotype is determined by comparing its pattern of reaction against the whole set of probes and not, as previously done in hybridisation assays, in an all-or-nothing basis. Interpretation of the results is objective and done through a visual basic application that compares the rates of hydrolysis of the ELISA substrate of an assayed isolate to a matrix of rates of hydrolysis obtained from standard haplotypes. This assay was validated and showed a better ability to resolve haplotypes than other assays to which it was compared experimentally. It may be automated to the same extent as any ELISA.

Item Type: Article
Uncontrolled Keywords: CTV, typing, CP phylogeny, stem-pitting, seedling yellows, quick decline, probe hybridisation
Subjects: Q Science > QH Natural history > QH301 Biology
Pre-2014 Departments: School of Science
Related URLs:
Last Modified: 14 Oct 2016 09:25
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/10305

Actions (login required)

View Item View Item