A single-tube reverse transcriptase-polymerase chain reaction (RT-PCR) using a fluorogenic real-time PCR detection method is described for the quantitation of equine infectious anemia virus (EIAV) RNA in the plasma of equids. To compensate for variations inherent in sample preparation a multiplex real-time RT-PCR system was developed that permitted the simultaneous calculation of the nucleic acid recovery rate along with the copy number of viral RNA molecules. Detection of EIAV RNA was linear from 109 to 101 molecules with intra- and inter-assay variability of less than 1% at 108, 106, 104 and 102 molecules.
|Number of pages||9|
|Journal||Journal of Virological Methods|
|State||Published - 2002|
Bibliographical noteFunding Information:
We thank Diane Furry for preparation of the manuscript. This work was supported by National Institutes of Health grants R01CA49296 and RO1A125850, by funds from the Lucille P. Markey Charitable Trust and the Kentucky Agricultural Experimental Station.
- Dual-labelled fluorogenic probes
- Quantitation of viral load
- Real-time PCR
ASJC Scopus subject areas