An improved in situ hybridization technique increased sensitivity at least 10-fold, allowing the detection and quantitation of 10-20 copies of visna virus RNA per cell after 2 days of exposure.
The sensitivity of in situ hybridization has been increased at least 10-fold by hybridizing in cDNA excess, by increasing the diffusion of the cDNA through the cells, by hybridizing at optimum temperature, and by stabilizing hybrids during autoradiography. Saturation of intracellular RNA with 3HcDNA has been achieved. The assay is quantitative. In situ hybridization has been used to detect and quantitate visna virus RNA in infected cells. By using 3HcDNA with specific activity of 2 X 10(8) dpm/micrograms and conditions that reduce background to negligible levels, 10--20 copies of viral RNA per cell can be detected and quantitated after 2 days of autoradiographic exposure.
Brahic et al. (Fri,) conducted a other in Visna virus infection (in vitro). Improved in situ hybridization technique vs. Standard in situ hybridization was evaluated on Detection sensitivity of viral RNA copies per cell. An improved in situ hybridization technique increased sensitivity at least 10-fold, allowing the detection and quantitation of 10-20 copies of visna virus RNA per cell after 2 days of exposure.