Detection of DNA viruses by radioactive and non radioactive DNA probes: Application to African Swine Fever virus
- 1 September 1987
- journal article
- research article
- Published by Springer Nature in Archiv für die gesamte Virusforschung
- Vol. 92 (3-4) , 233-242
- https://doi.org/10.1007/bf01317480
Abstract
A molecular hybridization technique using radioactive and non radioactive DNA probes, has been used to detect ASFV DNA immobilized on nitrocellulose paper. It is based on the use of plasmid pRPEL-2 as a hybridization probe. This plasmid containe the H-ClaI DNA fragment (size 5.6 Kbp) from the Spain-70 strain of ASFV. The sensitivity of detection using radioactive32P-probes (specific activity about 2×108 cpm per µg) was about 20 pg of viral DNA. The32P-pRPEL-2 DNA probe can detect about 100 infected MS cells and failed to hybridize to DNA from HSV-2, MS cells or salmon sperm. The sensitivity with non radioactive probes was about 4 ng of viral DNA for a sulfonated DNA probe and 400 pg for a biotinylated DNA probe. The effiency of DNA fixation to the filter, the effect of EDTA and of ultrasonic treatment of the sample were also investigated.This publication has 19 references indexed in Scilit:
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