Detection of human parvovirus B19 DNA by dot-hybridization and the polymerase chain reaction: applications for diagnosis of infections.
The development of the polymerase chain reaction for the detection of human parvovirus B19 DNA in clinical specimens is described. The sensitivity of this detection is compared with detection of virus using radiolabeled DNA and RNA probes. The polymerase chain reaction was the most sensitive, detecting approximately 10 fg B19 DNA. Several applications of the polymerase chain reaction for B19 DNA detection are discussed: i) Detection of B19 DNA sequences in paraffin-embedded fetal tissues in a case of intrauterine fetal death; ii) The presence of B19 DNA in a case of B19 infection after a bone marrow transplantation was demonstrated for a longer period of time as compared to detection with dot-hybridization; iii) In a considerable number of sera, proven to be B19 specific IgM positive, the presence of B19 DNA was demonstrated. The sensitivity of the polymerase chain reaction, combined with the simplicity, reduced time scale and range of applications, demonstrates the potential of this technique as an additional method for routine B19 diagnosis next to the detection of B19 specific antibodies.
Salimans
M M
MM
Central Clinical Virological Laboratory, University Hospital Leiden, The Netherlands.
D000914:Antibodies, Viral / Q000032:analysis; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D005313:Fetal Death; D005333:Fetus / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010321:Parvoviridae / Q000235:genetics / Q000302:isolation & purification*; D010322:Parvoviridae Infections / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D015183:Restriction Mapping
D000914:Antibodies, Viral; D004279:DNA, Viral; D015345:Oligonucleotide Probes