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Ultra-sensitive detection of two garlic potyviruses using a real-time fluorescent (Taqman®) RT-PCR assay
- Source :
-
Journal of Virological Methods . Jun2004, Vol. 118 Issue 1, p15-21. 7p. - Publication Year :
- 2004
-
Abstract
- A method for the detection of Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), the two most prevalent garlic potyviruses, has been developed that combines IC-RT-PCR/RT-PCR with the use of TaqMan® probes. Comparisons with ELISA results obtained with identical OYDV and LYSV infected samples showed sensitivity in detecting these viruses increased up to 106-fold. OYDV and LYSV were detected using different fluorochromes in the probe, thus allowing unequivocal diagnosis for each of them. The polyvalence of the designed virus-specific primers and probes was shown through their application to the detection of three isolates from very different geographical areas and from different hosts. A second version of the method avoids the need for an immunocapture step through the performance of a TaqMan® RT-PCR assay directly on extracts of garlic cloves. This modification on the proposed basic method allows the analysis of bulb samples in 3–4 h but did not give reproducible results with leaves. Both versions of the new diagnostic method bear great potential for their implementation in virus-free certification schemes in garlic, a vegetatively propagated crop for which such a certification is critical for a high-quality product. [Copyright &y& Elsevier]
- Subjects :
- *POTYVIRUSES
*SYMPTOMS
*FLUOROCARBONS
*PLANT viruses
Subjects
Details
- Language :
- English
- ISSN :
- 01660934
- Volume :
- 118
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Journal of Virological Methods
- Publication Type :
- Academic Journal
- Accession number :
- 13178960
- Full Text :
- https://doi.org/10.1016/j.jviromet.2004.01.012