Characterization of a nested polymerase chain reaction assay for detection of parvovirus B19.
The characterization and application of a nested polymerase chain reaction (PCR) assay for the detection of human parvovirus B19 DNA is described. The assay was evaluated with 149 diagnostic serum samples (collected up to 150 days after the onset of symptoms) previously tested by dot blot hybridization for B19 DNA and by class-specific capture radioimmunoassays for the detection of B19 immunoglobulin M (IgM) and IgG. B19 DNA was detectable by the PCR in 70% of the sera. There was a statistically significant association between the detection of B19 DNA by PCR and high B19 IgM values (P < 0.005), low B19 IgG values (P < 0.05), and a short interval between onset of symptoms and serum collection (P < 0.005). Serial serum samples, throat swabs, and peripheral blood mononuclear cells collected from 10 individuals during an outbreak of parvovirus B19 were also tested by the nested PCR. B19 DNA was detectable in the throat swabs at the time of the clinical illness and in the peripheral blood mononuclear cell fraction up to the end point of the study 6 months after infection. The location of the B19 DNA could not be determined in cytocentrifuge preparations of peripheral blood mononuclear cells with nonisotopic in situ hybridization and immunolabelling.
Patou
G
G
Department of Medical Microbiology, University College & Middlesex School of Medicine, London, United Kingdom.
Pillay
D
D
Myint
S
S
Pattison
J
J
D000914:Antibodies, Viral / Q000097:blood; D001483:Base Sequence; D001769:Blood / Q000382:microbiology; D004279:DNA, Viral / Q000097:blood*; D016731:Erythema Infectiosum / Q000175:diagnosis / Q000453:epidemiology; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D017403:In Situ Hybridization; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D008137:Longitudinal Studies; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D016732:Parvovirus B19, Human / Q000235:genetics / Q000302:isolation & purification*; D010614:Pharynx / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011863:Radioimmunoassay; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D004279:DNA, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M