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ARS Home » Research » Publications at this Location » Publication #72027


item Figueira, Antonia
item Domier, Leslie
item D'arcy, Cleora

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/5/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary: Barley yellow dwarf viruses infects cereal crops causing significant economic losses each year. Several different methods have been developed to detect the presence of viruses in infected plants. The sensitivities of three different techniques were evaluated in this study. The first technique used antibodies to detect proteins produced by the viruses. This technique is the most commonly used technique for the detection of plant viruses and is relatively simple and effective. The second and third techniques detected the RNA, or genetic material, of barley yellow dwarf viruses. The second technique, "nucleic acid hybridization", was as effective and sensitive as standard antibody techniques. However, it was somewhat more complex than antibody-based methods would be, an effective alternative only when antibodies that recognize the viruses are not available. The third technique, "polymerase chain reaction", was about 1000 times more sensitive than either of the first two techniques. The polymerase chain reaction assay would be the best method by which to detect barley yellow dwarf viruses in situations where the greatest sensitivity was requied. This paper directly compared three commonly used techniques for virus detection based on sensitivity and ease of use. The information provided will be useful to scientists who are interested in the relative merits of these techniques.

Technical Abstract: Detection of barley yellow dwarf virus (BYDV-PAV-IL) by an improved nucleic acid hybridization technique, using a nonradioactive probe with chromogenic and chemiluminescent substrates, was compared to detection by PCR, double antibody sandwich ELISA with polyclonal antibodies, and triple antibody sandwich ELISA with polyclonal and monoclonal antibodies. Each method was used for detection of purified virus and virus in sap extracts from infected oat leaves. The detection limits for both ELISA procedures were 1 ng of purified BYDV-PAV-IL and the equivalent of 78 ng of infected tissue. Nucleic acid hybridization with either chemiluminescent or chromogenic substrates also detected down to 1 ng of purified BYDV-PAV-IL, but was slightly more sensitive for detection of virus in tissue extracts (25 ng of infected tissue). The most sensitive detection technique was PCR amplification, which could detect as little as 0.1 pg of RNA extracted from purified virus and detected viral RNA in the equivalent of 0.5 pg of infected leaf tissue.