Detection of parvovirus DNA in human serum using biotinylated RNA hybridisation probes.
Methods were established for the detection of parvovirus DNA in human serum using single-stranded RNA probes. The sensitivity of detection of virus using 32P-radiolabelled RNA versus non-radiolabelled biotinylated RNA probes using a streptavidin-polyalkaline phosphatase detection system was compared. Virus was detected using 32P-labelled and biotinylated RNA probes at serum dilutions of 10(-3), equivalent to approx. 3 pg of viral DNA. Using biotinylated RNA probes and a dot-blot system, diagnosis of numerous serum samples could be performed within 8 h of receipt of samples, using an RNA probe which was synthesised and stored at -20 degrees C for up to 12 months without loss of sensitivity. Our work demonstrates the potential of biotinylated RNA probes in the routine analysis of viral sequences in serum.
Cunningham
D A
DA
Department of Biochemistry, University College, London, U.K.
Pattison
J R
JR
Craig
R K
RK
D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D009693:Nucleic Acid Hybridization; D010321:Parvoviridae / Q000302:isolation & purification*; D010322:Parvoviridae Infections / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D012367:RNA, Viral
D004279:DNA, Viral; D012367:RNA, Viral