Graduate Thesis Or Dissertation

 

The detection of ilarvirus (subgroup B) by enzyme-linked immunosorbent assay Public Deposited

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  • The reliability and sensitivity of enzyme-linked immunosorbent assay (ELISA) in the detection of ilarvirus (isometric labile ringspot virus) subgroup B and prune dwarf virus (PDV) isolates in Prunus and Malus species has been well established. To use this technique in large scale virus indexing programs more information was required on the reactions of commercially available antisera against the wide range of naturally occurring ilarvirus isolates. Specifically, information was required to determine whether an antiserum, or a group of antisera, exists that will detect the entire spectrum of ilarvirus subgroup B and PDV isolates (here-after referred to as ILAR-b type isolates) in ELISA tests. In this thesis 44 isolates of ILAR-b have been tested by ELISA using eleven antisera prepared against isolates of this group. The virus isolates used were collected by previous workers in 12 states and four countries from a multitude of hosts, and cover a wide range of the isolates originally referred to by R. W. Fulton (1968) as ILAR viruses. Six of the antisera used were commercially available. Only three of the 44 isolates reported to be the ILAR-b type were not detected by any of the eleven antisera used. Those isolates detected by ELISA fell into three of the serogroups originally described by Fulton for this type of virus, namely a necrotic ringspot virus (NRSV) serogroup, an apple mosaic virus (ApMV) serogroup, and a prune dwarf virus (PDV) serogroup. Other serogroups described for ilarviruses were not used in this research. In these ELISA tests, the ability to detect a virus isolate in one serogroup using antisera of another serogroup was quite poor, whereas within-group detection was fairly uniform for all isolate-antiserum combinations. A mixture of the best antisera of each serogroup was able to detect all isolates previously detected individually by the three separate antisera. Bioassays employing Chenopodium quinoa and Cucumis sativus, detected ILAR-b isolates in young spring tissue of Malus and Prunus sp. with reliability about equal to ELISA. ELISA, however, was far more efficient (space, material, and time-wise) and proved more sensitive over an extended period of the growing season. It is hoped that the research results presented will: 1. Document those strains of ILAR-b type viruses that can be detected by ELISA using the presently available antisera. 2. Facilitate increased use of ELISA in virus certification programs.
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