Comparison of nonradioactive cDNA probes for detection of potato spindle tuber viroid by dot-blot hybridization assay

Studies on the diagnosis of hop stunt viroid in fruit trees: Identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents

“…Regarding sensitivity, the nearly full-length probe described here performs as well as previous radioactive or multimeric probes described for other viroids (Roy et al, 1989;Candresse et al, 1990;Kanematsu et al, 1991;Singh et al, 1994), clearly detecting subpicogram amounts of HSVd RNAs. HSVd was also detected in dilutions of infected plant extracts corresponding to 12.5 ng of analysed tissue (Figure lb).…”

“…A further improvement arose with the advent of non-radioactive methods for the detection of hybridized probes, allowing their progressive introduction as tools in the study of plant viruses (Habili et al, 1987;Eweida etal., 1990;Fouly et al, 1992;Gemmrich etal., 1993;M~s et al, 1993;Dietzgen etal., 1994;Harper and Creamer, 1995;S~nchez-Navarro et al, 1996). Several reports have already described the use of these non-radioactive probes to detect viroid RNAs, albeit most of them correspond to potato spindle tuber viroid (PSTVd) (McInnes et al, 1989;Roy et al, 1989;Candresse et al, 1990;Hopp et al, 1991;Kanematsu et al, 1991;Welnicki and Hiruki, 1992;Podleckis et al, 1993;Singh et al, 1994). Recently, a non-radioactive tissue-print hybridization method was reported for viroid detection (Romero-Durb~n et al, 1995), although the authors encountered problems in the detection of HSVd.…”

Nucleic Acid Hybridization

“…While it is feasible on paper to evaluate your research needs against these criteria, the decision must usually be determined at the lab bench. In lieu of the need to test individual systems, several studies compared the sensitivity of nonradioactive probes to the 32 P gold standard: Yang et al (1999), Plath, Peters, and Einspanier (1996), Nass and Dickson (1995), Moore and Margolin (1993), Puchhammer-Stoeckl, Heinz, and Kunz (1993), Engler-Blum et al (1993, Bright et al (1992), Kanematsu et al (1991), Lion and Haas (1990), Jiang, Estes, and Metcalf (1987), Tenberge et al (1998), Holtke et al (1992), Pollard-Knight et al (1990), Hill and Crampton (1994), Dubitsky, Brown, andBrandwein (1992), andNakagami et al (1991).…”