@ARTICLE{TreeBASE2Ref2110,
author = {Filomena Fonseca and Gustavo Nolasco and Carla Santos and Gon?alo Silva},
title = {Development of an asymmetric PCR ELISA typing assay for citrus tristeza virus based on the coat protein gene.},
year = {2008},
keywords = {},
doi = {},
url = {},
pmid = {},
journal = {Journal of Virological Methods},
volume = {},
number = {},
pages = {},
abstract = {The coat protein gene of isolates of citrus tristeza virus (CTV) from twenty 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 hybridization 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 hybridization 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 experimentally compared. It may be automated to the same extent as any ELISA assay.}
}
Citation for Study 2173

Citation title:
"Development of an asymmetric PCR ELISA typing assay for citrus tristeza virus based on the coat protein gene.".

This study was previously identified under the legacy study ID S2179
(Status: Published).
Citation
Fonseca F., Nolasco G., Santos C., & Silva G. 2008. Development of an asymmetric PCR ELISA typing assay for citrus tristeza virus based on the coat protein gene. Journal of Virological Methods, null.
Authors
-
Fonseca F.
-
Nolasco G.
-
Santos C.
-
Silva G.
Abstract
The coat protein gene of isolates of citrus tristeza virus (CTV) from twenty 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 hybridization 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 hybridization 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 experimentally compared. It may be automated to the same extent as any ELISA assay.
About this resource
- Canonical resource URI:
http://purl.org/phylo/treebase/phylows/study/TB2:S2173
- Other versions:
Nexus
NeXML
- Show BibTeX reference
@ARTICLE{TreeBASE2Ref2110,
author = {Filomena Fonseca and Gustavo Nolasco and Carla Santos and Gon?alo Silva},
title = {Development of an asymmetric PCR ELISA typing assay for citrus tristeza virus based on the coat protein gene.},
year = {2008},
keywords = {},
doi = {},
url = {},
pmid = {},
journal = {Journal of Virological Methods},
volume = {},
number = {},
pages = {},
abstract = {The coat protein gene of isolates of citrus tristeza virus (CTV) from twenty 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 hybridization 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 hybridization 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 experimentally compared. It may be automated to the same extent as any ELISA assay.}
}
- Show RIS reference
TY - JOUR
ID - 2110
AU - Fonseca,Filomena
AU - Nolasco,Gustavo
AU - Santos,Carla
AU - Silva,Gon?alo
T1 - Development of an asymmetric PCR ELISA typing assay for citrus tristeza virus based on the coat protein gene.
PY - 2008
KW -
UR -
N2 - The coat protein gene of isolates of citrus tristeza virus (CTV) from twenty 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 hybridization 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 hybridization 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 experimentally compared. It may be automated to the same extent as any ELISA assay.
L3 -
JF - Journal of Virological Methods
VL -
IS -
ER -