Serological Detection of Members of the Potyviridae with Polyclonal Antisera

Abstract

The cross‐reactivity of two polyclonal antisera with various members of the Potyviridae was determined using four different serological methods. An antiserum (no. 314) prepared against apparently intact turnip mosaic potyvirus (TuMV) particles showed a high cross‐reactivity with all of 20 aphid‐borne potyviruses tested as well as the mite‐borne ryegrass mosaic rymovirus (RGMV) and the fungus‐borne barley mild mosaic bymovirus (BaMMV) with two of the four methods used (indirect PTA‐ELISA or IEM and Western blotting). In immunoelectron microscopic decoration, about two thirds of the viruses tested reacted positively whereas in the direct DAS‐ELISA only the homologous TuMV reacted positively. Another antiserum (no. 203) produced by using as immunogen a mixture of the dissociated core proteins of three trypsin‐treated potyviruses (potato A and V and plum pox viruses), reacted positively in both indirect ELISA and Western blotting with all aphid‐borne potyviruses and RGMV. No reactions were obtained with the other two methods. The causes for these different reactivities are discussed. Both antisera are useful for the detection of aphid‐borne potyviruses, but one (no. 314) is especially suitable for the detection of these viruses under routine conditions by use of a simple indirect PTA‐ELISA.

Sharka: The Past, The Present and The Future

“…ELISA method underwent progress with some modifications and now, it is one of the most popular and used techniques for PPV detection [359,360,361,362]. Whereas the usage of polyclonal antibodies, which recognize multiple epitopes on any one antigen, is controversial due to problems with specificity and consequently with sensitivity (serum contains a mixture of antibodies of different affinity), monoclonal antibodies, which detect only one epitope on the antigen, helped to overcome above-mentioned problems and are still widely used in PPV diagnostics [363,364,365,366,367]. Monoclonal antibodies are usually obtained after immunization of BALB/c mice with purified PPV isolates [367,368].…”

Evidence of a plant virus in Iceland

“…This supposition was confirmed by means of plate trapped antigen (PTA-ELISA) using a potyvirus group-specific polyclonal antiserum (Richter et al, 1994) or the monoclonal antibody 3H8 (Richter et al, 1995).…”

“…Virus identification was carried out by using the immunoelectron-microscopical method of decoration with antisera (Milne and Luisoni, 1977 For serological investigations standard DAS-ELISA as described by Clark and Adams (1977) and the plate trapped antigen (PTA)-ELISA according to Richter et al (1994) were carried out. Sample preparation and SDS-PAGF for "Western blotting were used as described by Laemmli (1970).…”

Sequence of the 3′-terminal region of the RNA of a mite transmitted potyvirus fromHordeum murinum L.

“…A polyclonal reference antiserum against WSMV-H81 was provided by S. A. Lommel (Raleigh). For some comparative investigations a polyclonal antiserum produced against turnip mosaic virus (TuMV-314) displaying a high degree of cross-reactivity with different members of aphid-borne potyviruses [Richter et al, 1994] was included.…”