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Updated: Aug 11, 2026
Efficient Recombinant Parvovirus Production with the Help of Adenovirus-derived Systems
Published on: April 23, 2012
Development of an improved method of detection of infectious parvovirus B19
1Hematology Branch, National Heart, Lung and Blood Institute, Bethesda, MD, USA.
Background:
Parvovirus B19, the only known pathogenic human parvovirus is the aetiologic agent of erythema infectiosum, transient aplastic crisis, pure red cell aplasia, and hydrops fetalis. Transmission is either by respiratory secretions or, as it can be present at high titre in plasma, by blood and blood products. B19 is only cultured with difficulty in vitro, and there is no readily available assay for detecting B19 infectivity or neutralizing antibodies.
Objectives:
In this study, we evaluated different methods to detect viral infection for the purpose of developing automated methods for large-scale testing of viral infectivity, development of neutralizing antibody and viral inactivation assays.
Study Design:
Different cell lines were evaluated for their ability to support B19 infection and assays tested for sensitivity and ease of performing. A high-throughput assay was validated by determining infectious virus in blood pools and for determining neutralizing antibody in sera.
Results:
B19 protein production was detected by immunofluorescence (IF) staining and increased viral DNA production by dot blot hybridization and quantitative PCR. The detection of RNA transcripts by RT-PCR assay and quantitative RT-PCR (qRT-PCR) was used as an indirect marker for infection. Of the cell lines tested, the subclone UT7/Epo-S1 showed the greatest sensitivity to B19 infection, with detection of viral transcripts by qRT-PCR the preferred assay. The assays were validated by experiments to determine the infectious titre of sera from acutely infected humans, to evaluate the presence of infectious virus in human donor plasma pools and to measure neutralizing antibodies.
Insights
Developing sensitive assays for Parvovirus B19 (B19) infection is crucial for blood safety. Quantitative RT-PCR on UT7/Epo-S1 cells is the preferred method for detecting B19 viral transcripts and neutralizing antibodies.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Parvovirus B19 (B19) is a significant human pathogen causing various clinical conditions.
- B19 transmission occurs via respiratory secretions and blood products.
- Current methods for detecting B19 infectivity and neutralizing antibodies are limited.
Purpose of the Study:
- To evaluate different methods for detecting B19 viral infection.
- To develop automated, high-throughput assays for large-scale B19 testing.
- To establish assays for quantifying neutralizing antibodies and viral inactivation.
Main Methods:
- Screening of various cell lines for B19 susceptibility.
- Assessing assay sensitivity and ease of performance.
- Validation of high-throughput assays using blood pools and human sera.
Main Results:
- UT7/Epo-S1 cell subclone demonstrated highest sensitivity to B19 infection.
- Quantitative RT-PCR (qRT-PCR) effectively detected B19 RNA transcripts.
- Assays were validated for determining infectious B19 titers and neutralizing antibodies.
Conclusions:
- Quantitative RT-PCR on UT7/Epo-S1 cells is a preferred method for B19 detection.
- Developed assays facilitate large-scale testing of B19 infectivity and antibody responses.
- These advancements contribute to improved blood product safety and B19 diagnostics.
