Infection of the erythroid cell line, KU812Ep6 with human parvovirus B19 and its application to titration of B19 infectivity

Infection of the erythroid cell line, KU812Ep6 with human parvovirus B19 and its application to... 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 TCID 50 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 TCID 50 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 TCID 50 ml to <10 TCID 50 ml (lower limit of detection) at 60°C for 3 h or at 70°C for 30 min, but only decreased to 10 2.5 TCID 50 ml at 50°C for 8 h. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Virological Methods Elsevier

Infection of the erythroid cell line, KU812Ep6 with human parvovirus B19 and its application to titration of B19 infectivity

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Publisher
Elsevier
Copyright
Copyright © 1999 Elsevier Science B.V.
ISSN
0166-0934
eISSN
1879-0984
D.O.I.
10.1016/S0166-0934(99)00105-6
Publisher site
See Article on Publisher Site

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 TCID 50 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 TCID 50 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 TCID 50 ml to <10 TCID 50 ml (lower limit of detection) at 60°C for 3 h or at 70°C for 30 min, but only decreased to 10 2.5 TCID 50 ml at 50°C for 8 h.

Journal

Journal of Virological MethodsElsevier

Published: Dec 1, 1999

References

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