Quick Characterization of Citrus Tristeza Virus Isolates by Capillary Electrophoresis-Single-Strand Conformation Polymorphism

Raspagliesi, D.; Licciardello, Grazia; Lombardo, Giorgio; Catara, A.; Rizza, S.

doi:10.17660/actahortic.2011.892.22

Cited by 2 publications

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“…Five of the isolates, classified as biotype 4 (severe), were obtained from local sweet orange trees grafted on sour orange: isolates SG29, S24 and S25 were from a 25-yr-old declining 'Sanguinello' sweet orange (7), while isolate S9 was from a declining "Tarocco comune" sweet orange and isolate S13 from a "Tarocco Gallo" sweet orange. Three isolates belonged to biotype 2 (mild): isolate TDV was collected from a 5-yr-old symptomless 'Tarocco' sweet orange tree grafted on Troyer citrange (19), M1 from a sour orange seedling naturally infected by aphids and M2 from a "Tarocco Sant'Alfio" grafted on sour orange.…”

Section: Source Of Virus Isolatesmentioning

confidence: 99%

“…In plant pathology, CE-SSCP has recently been applied for screening genetic diversity in a Plum pox virus population (8). In a previous study, we reported the use of CE-SSCP for the characterization of CTV isolates using the Applied Biosystem 3130 capillary electrophoresis apparatus in "non denaturing" conditions (19), modifying the running protocol which had previously only been calibrated for human pathology studies (14).…”

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confidence: 99%

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A New Approach for the Rapid Detection and Genetic Discrimination of Mild and Severe Isolates of Citrus tristeza virus Based on ELISA/CE-SSCP

Raspagliesi¹,

Licciardello²,

Lombardo³

et al. 2011

International Organization of Citrus Virologists Conference Proceedings (1957-2010)

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A new approach for the rapid discrimination of mild and severe Citrus tristeza virus (CTV) isolates was developed by a sequential process made up of the DAS-ELISA immunological test followed by fluorescence-based Capillary Electrophoresis-Single Strand Conformation Polymorphism (CE-SSCP) analysis. The new method helps both in locating CTV infected trees and in preliminarily typing of the virus isolates, thus saving both time and resources. Partial p18 gene amplification products of CTV-RNAs directly extracted from infected leaves or recovered from ELISA plates are presented. Specific profiles of forward and reverse strands were obtained when biologically distinct CTV isolates were analysed both directly and following ELISA. The CE-SSCP method is simple, highly sensitive and highly reproducible, and can process a large number of samples for a variety of needs. Applied in a combined process, ELISA and CE-SSCP helped simultaneously to detect the virus and to collect useful information on the genetic diversity and structure of the CTV population present in a territory.

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Section: Source Of Virus Isolatesmentioning

confidence: 99%

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confidence: 99%