Detection of B19 parvovirus infections by a dot-blot hybridization assay using a digoxigenin-labelled probe.
A non-radioactive dot-blot hybridization assay for the detection of B19 parvovirus infections was developed using a digoxigenin-labelled probe both on nylon and nitrocellulose filters. A 700 bp BamHI HindIII fragment of B19 DNA was used to construct the probe. Probe labelling was carried out by incorporating deoxyuridine triphosphate labelled with digoxigenin. The dot-blot hybridization assay was visualized by an immunoenzymatic reaction using antidigoxigenin Fab fragments labelled with alkaline phosphatase. The specificity and sensitivity of digoxigenin-labelled B19 DNA probe was compared with the results obtained with 32P-labelled B19 DNA probe. Out of the 504 serum samples tested, 3 samples were positive in all the hybridization assays performed and 494 were negative, 7 serum samples gave a weak positive reaction when Dig-B19 probe was used on nitrocellulose filters. The 77 pharyngeal swabs tested were negative in all the hybridization assays performed. Our hybridization assay showed a high sensitivity and reproducibility and it appears to be a rapid, practical and reliable test for routine screening of B19 parvovirus DNA in large numbers of clinical specimens.
Azzi
A
A
Institute of Microbiology, University of Firenze, Italy.
Zakrzewska
K
K
Gentilomi
G
G
Musiani
M
M
Zerbini
M
M
D000293:Adolescent; D002648:Child; D003101:Collodion; D015342:DNA Probes; D004279:DNA, Viral / Q000302:isolation & purification*; D004076:Digoxigenin; D006801:Humans; D009693:Nucleic Acid Hybridization; D009757:Nylons; D010321:Parvoviridae / Q000302:isolation & purification; D010322:Parvoviridae Infections / Q000175:diagnosis* / Q000382:microbiology; D014773:Virology / Q000379:methods
D015342:DNA Probes; D004279:DNA, Viral; D009757:Nylons; D003101:Collodion; D004076:Digoxigenin