Detection and characterization of Plum pox virus: serological methods

Abstract

Tremendous progress has been made in the research and development of Plum pox virus (PPV) serological reagents and methods in recent years. Two facts have revolutionised the serological detection and characterization of the virus: the development of the ELISA method in 1977, and the later emergence of specific monoclonal antibody technology. The availability of commercial kits has popularised PPV diagnosis, now making diagnosis possible at large scale for quarantine purposes, eradication programmes and control of the disease in nurseries. The use of the universal monoclonal antibody 5B-IVIA, used in DASI-ELISA, is the most accurate system for routine PPV detection. Likewise, the use of typing monoclonal antibodies gives exact characterization of the main PPV types described: 4DG5 for PPV-D, AL for PPV-M, EA24 for PPV-EA, and TUV and AC for PPV-C. There is, in general, an excellent correlation between serological data obtained with PPV specific monoclonal antibodies and data obtained by molecular PCR based methods. ELISA using a single or a mixture of monoclonal antibodies will remain the preferred method for universal detection and routine screening of PPV for years to come. Today, other serological methods and reagents are also recommended in the EPPO Diagnostic Protocol, increasing the number of reliable tests available for PPV detection. These developments have helped to control sharka disease in recent years. International co-operation in this field has been crucial to the improvement and validation of serological tools for PPV detection and characterization.

PPV susceptibility of commonly used peach rootstock-scion combinations

“…Th ree strains (PPV-M, PPV-D and PPV-Rec) are widely spread (Dallot et al, 1998;Myrta et al, 1998;Glasa et al, 2004), but six have specifi c geographical locations or infrequent occurrence (PPV-EA, PPV-C, PPV-W, PPV-T, PPV-Man, PPV-CR; Candresse et al, 1994;Nemchinov and Hadidi, 1996;James et al, 2003;Serçe et al, 2009;Palmisano et al, 2012;Chirkov et al, 2013;Glasa et al, 2013). Sharka disease heavily determines the world's stone fruit production, and causes loss of Euro millions every year (Cambra et al, 2006). PPV infects not only stone fruits but almond (Pribék et al, 2001) and blackthorn (Salamon and Palkovics, 2002), thus a natural wild host species endanger orchards as a reservoir and source of the virus infection by aphid transmission.…”

Development of Latex Agglutination Test for Rapid Detection of Citrus Tristeza Closterovirus

“…Eradication programs and the use of cross-protection strategies with mild isolates can also contribute to the control of the disease (Roistacher and Moreno, 1991). All these control strategies require a large number of detection tests usually performed by enzymelinked immunesorbent assays (Clark et al, 1976;Clark and Adams, 1977) and PCR-based assays (Cambra et al, 2006). This report describes the development of latex agglutination test (LAT) as rapid, sensitive and reliable method and its application in CTV monitoring in citrus plants.…”

Epidemiología de Plum pox virus y Citrus tristeza virus en bloques de plantas de vivero. Métodos de control

“…Existen varios tipos de aislados de este virus, que varían en su sintomatología, epidemiología y agresividad. Estas diferencias han sido serológica y molecularmente descritas (Candresse y Cambra et al, 2006a) De todos ellos, los tipos más predominantes son PPV-D y PPV-M. El tipo PPV-D, único dispersado ampliamente en España, causa daños en albaricoquero y en ciruelo europeo, produciendo graves pérdidas en variedades tempranas de los mismos, pero afecta poco a ciruelo japonés, y no se dispersa naturalmente entre cultivares de melocotonero o lo hace con poca eficacia. No infecta a almendros (P. dulcis) y cerezos (P. cerasus).…”

“…Este anticuerpo es el único que puede detectar todos los aislados de PPV citados hasta la fecha (Cambra et al, 2006a). En cuanto a la caracterización de los aislados, el mismo protocolo indica que para la caracterización de los aislados PPV-D recomienda los anticuerpos monoclonales 4DG5 y 4DG11 (Cambra et al, 1994) y para la caracterización de aislados PPV-M recomienda el anticuerpo monoclonal AL 1.…”

“…Introducción 15 (Boscia et al, 1997). Este anticuerpo también reconoce a aislados del tipo Rec (Cambra et al, 2006a). Otros anticuerpos monoclonales útiles en la caracterización de aislados de PPV son: EA24 (Myrta et al, 1998) que detecta aislados del tipo El Amar y los anticuerpos AC y TUV (Myrta et al, 2000), que detectan aislados del tipo PPV-C. El uso de anticuerpos policlonales no está recomendado debido a su pobre especificidad, baja homogeneidad entre las partidas y que sólo del 10-15% de los anticuerpos obtenidos reaccionan contra el antígeno diana, además se pueden dar fenómenos de reacción cruzada entre el anticuerpo policlonal y otros virus o huéspedes (Cambra et al, 2006a).…”