TY - JOUR
T1 - Evaluation of anti-parvovirus B19 activity in sera by assay using quantitative polymerase chain reaction
AU - Saito, Takako
AU - Munakata, Yasuhiko
AU - Fu, Yi
AU - Fujii, Hiroshi
AU - Kodera, Takao
AU - Miyagawa, Eiji
AU - Ishii, Keiko
AU - Sasaki, Takeshi
N1 - Funding Information:
This work was supported by grant-in aid for scientific research (A) from the Ministry of Education, Science, Sports and Culture in Japan, and by grants to T.S. awarded from the Kurozumi Medical Foundation. Informed consent was obtained from all patients in this study.
PY - 2003/1
Y1 - 2003/1
N2 - 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.
AB - 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.
KW - Human parvovirus B19
KW - KU812Ep6
KW - Neutralizing antibody
KW - Quantitative PCR
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U2 - 10.1016/S0166-0934(02)00195-7
DO - 10.1016/S0166-0934(02)00195-7
M3 - Article
C2 - 12445941
AN - SCOPUS:0037210837
VL - 107
SP - 81
EP - 87
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -