Summary
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.
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Tabarés, E. Detection of DNA viruses by radioactive and non radioactive DNA probes: Application to African Swine Fever virus. Archives of Virology 92, 233–242 (1987). https://doi.org/10.1007/BF01317480
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DOI: https://doi.org/10.1007/BF01317480