RRC ID 39363
著者 Miyagawa E, Yoshida T, Takahashi H, Yamaguchi K, Nagano T, Kiriyama Y, Okochi K, Sato H.
タイトル Infection of the erythroid cell line, KU812Ep6 with human parvovirus B19 and its application to titration of B19 infectivity.
ジャーナル J Virol Methods
Abstract A human parvovirus B19 (B19) infectivity assay was developed using the erythroid cell line, KU812Ep6. KU812Ep6 was cloned for high efficiency infection with B19 in vitro, in the presence of erythropoietin by a limiting dilution method from the parent cell line, KU812. B19 was effectively propagated in KU812Ep6 and was detected for B19 antigens, VP1 and VP2. The titers of B19 positive sera measured with KU812Ep6 cells were in the range of 10(6) to 10(8) TCID50 ml. This KU812Ep6 infectivity assay had a 10(3)-10(4.5) higher sensitivity than the colony forming unit-erythroid (CFU-e) injury assay. It was calculated that one TCID50 needed 10(3) B19 genome copies, judging from the infectivity assay and semi-quantitative PCR. The KU812Ep6 infectivity assay was also used to determine infectivity of B19 in vitro, and to evaluate inactivation, as well as clearance of the virus. The inactivation of B19 by heating was carried out and infectivity declined from 10(4) TCID50 ml to < 10 TCID50 ml (lower limit of detection) at 60 degrees C for 3 h or at 70 degrees C for 30 min, but only decreased to 10(2.5) TCID50 ml at 50 degrees C for 8 h.
巻・号 83(1-2)
ページ 45-54
公開日 1999-12-1
DOI 10.1016/s0166-0934(99)00105-6
PII S0166-0934(99)00105-6
PMID 10598082
MeSH Antigens, Surface Base Sequence Cell Line Colony-Forming Units Assay DNA Primers / genetics Erythrocytes / immunology Erythrocytes / ultrastructure Erythrocytes / virology* Evaluation Studies as Topic Hot Temperature Humans Microscopy, Electron Parvovirus B19, Human / genetics Parvovirus B19, Human / pathogenicity* Parvovirus B19, Human / ultrastructure Polymerase Chain Reaction Virology / methods* Virulence
IF 1.786
引用数 59
WOS 分野 BIOCHEMICAL RESEARCH METHODS BIOTECHNOLOGY & APPLIED MICROBIOLOGY VIROLOGY
リソース情報
ヒト・動物細胞 KU812(RCB0495)