Evaluation of anti-parvovirus B19 activity in sera by assay using quantitative polymerase chain reaction.
Human parvovirus B19 (B19) infects cells of erythroid lineage. Production of neutralizing antibodies (Abs) is indispensable for recovery from B19-related disease state. In this study, we used a convenient method to measure neutralizing activities in human sera by using a real-time quantitative PCR based assay. Erythroid cell line KU812Ep6 was incubated with test sera before infection with B19 virus. The copy number of B19-DNA in cultures was decreased in the presence of the sera from patients who recovered from acute B19 infection, whereas no decrease in B19-DNA was in cultures incubated with sera from healthy volunteers who had no B19 infection. The decrease in B19-DNA copy number was calculated and the inhibition percentage was expressed as neutralizing activity to B19. A clinical study showed that the levels of neutralizing ability were high in patients who recovered soon after acute B19 infection, but were low in some patients with a prolonged clinical course for recovery from B19 infection. This method is simple and convenient compared with methods described previously, showing its usefulness to evaluate the neutralizing activity to B19.
Saito
Takako
T
Department of Molecular Diagnostics, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan.
Munakata
Yasuhiko
Y
Fu
Yi
Y
Fujii
Hiroshi
H
Kodera
Takao
T
Miyagawa
Eiji
E
Ishii
Keiko
K
Sasaki
Takeshi
T
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D016732:Parvovirus B19, Human / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods*
D000914:Antibodies, Viral