Back to Search
Start Over
Detection of citrus psorosis-ringspot virus using RT-PCR and DAS-ELISA.
- Source :
-
Plant Pathology . Dec97, Vol. 46 Issue 6, p830-836. 7p. - Publication Year :
- 1997
-
Abstract
- Psorosis, sometimes also associated with ringspot symptoms, is a widespread and damaging disease of citrus in many parts of the world including South America and the Mediterranean basin. We describe the application of RT-PCR and DAS-ELISA diagnostics to an isolate of citrus ringspot virus (CtRSV-4) and other virus isolates associated with this disease. Fragments of cDNA from bottom-component RNA of CtRSV-4 were cloned and sequenced, and PCR primers were designed, 5'ACAATAAGCAAGACAAC upstream, and 5 CCATGTCACTTCTATTC downstream. RT-PCR experiments using these primers allowed detection of CtRSV-4 in infected citrus leaves down to a tissue dilution of 1/12 800 representing 2μg of tissue, and less sensitive detection of the related citrus psorosis-associated vims (CPsAV90-l-l) and four other psorosis isolates from Argentina and the USA. In addition, CtRSV-4 particles were partially purified from local lesions in Chenopodium quinoa, and the preparations used to taise a rabbit antiserum. The antiserum was absorbed with extracts of healthy C quinoa leaves, and a DAS-ELISA kit was prepared and tested for detection of CtRSV-4, CPsAV90-l-l, and other psorosis isolates from Argentina, the USA, Italy and Spain. The ELISA detected CtRSV-4 down to a tissue dilution of 1/1600, and most other psorosis isolates down to dilutions of 1/200-1/800. Three of a total of 20 heterologous isolates were consistently negative. Comparison of the PCR and ELISA results suggests that both methods can be used for detection of a range of psorosis isolates, but that variation of the viruses in the field might cause problems for any one diagnostic test. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00320862
- Volume :
- 46
- Issue :
- 6
- Database :
- Academic Search Index
- Journal :
- Plant Pathology
- Publication Type :
- Academic Journal
- Accession number :
- 14409408
- Full Text :
- https://doi.org/10.1046/j.1365-3059.1997.d01-81.x