| RRC ID |
39363
|
| Author |
Miyagawa E, Yoshida T, Takahashi H, Yamaguchi K, Nagano T, Kiriyama Y, Okochi K, Sato H.
|
| Title |
Infection of the erythroid cell line, KU812Ep6 with human parvovirus B19 and its application to titration of B19 infectivity.
|
| Journal |
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.
|
| Volume |
83(1-2)
|
| Pages |
45-54
|
| Published |
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
|
| Times Cited |
59
|
|
WOS Category
|
BIOCHEMICAL RESEARCH METHODS
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
VIROLOGY
|
| Resource |
| Human and Animal Cells |
KU812(RCB0495) |
