Analysis of B19 virus contamination in plasma pools for manufacturing, by using a competitive polymerase chain reaction assay

Gallinella, Giorgio; Moretti, Elena; Nardi, G; Zuffi, Elisa; Bonvicini, Francesca; Bucci, E; Musiani, Monica; Zerbini, Marialuisa

doi:10.1046/j.1423-0410.2002.00227.x

Cited by 12 publications

(12 citation statements)

References 41 publications

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“…Twenty plasma pools, examined previously in a pilot study for the presence of B19 DNA [Gallinella et al, 2002], were investigated to determine the viral load, the infectivity and expression of B19 virus in KU812Ep6 cells and the level of anti-B19 antibodies.…”

Section: Resultsmentioning

confidence: 99%

“…Of the 20 plasma pools, nine were positive and 11 negative for B19 DNA [Gallinella et al, 2002]. To establish B19 viral load in nine positive plasma pools a titration was carried out by competitive PCR assay.…”

Section: Resultsmentioning

confidence: 99%

“…The 20 plasma pools had been screened previously in a pilot study by PCR-ELISA for the presence of B19 virus DNA. Of the 20 plasma pools, nine tested positive, and 11 negative for B19 DNA [Gallinella et al, 2002].…”

Section: Samplesmentioning

confidence: 99%

“…All 20 plasma pools were processed before amplification by using the AmpliScreen Lysis treatment (Roche Molecular Biochemicals, Roche Diagnostics S.p.A, Monza, Italy), according to the manufacturer's protocol, as described by Gallinella et al [2002].…”

Section: B19 Dna Titration By Competitive Pcr Assaymentioning

confidence: 99%

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Molecular testing for detection of in vitro infectivity of plasma pools contaminated with B19 virus

Bonvicini

Gallinella

Cricca

et al. 2004

Journal of Medical Virology

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B19 virus can be transmitted by contaminated blood or blood products. Recent observations, in healthy volunteers, suggest that active B19 infection can follow the administration of plasma pools with a concentration > or =10(7) genome equivalents/ml (geq/ml) of B19 DNA. However, patients receiving batches with levels of virus DNA lower than 10(4) geq/ml do not show any evidence of transmission of the virus. The aim of the study was to show, by in vitro assays, a threshold of viral load in B19 contaminated plasma pools over which the infection can be transmitted. Twenty plasma pools, each containing 960 single donations, were tested to correlate the viral load and the level of antibodies anti-B19 with the in vitro infectivity and expression of B19 virus. All the plasma pools, titrated for B19 viral load by competitive PCR, were inoculated into KU812Ep6 erythroid human cell line. Five of the nine contaminated plasma pools, with a B19 DNA concentration > or =3.60 x 10(6) geq/ml, were able to infect KU812Ep6 cells. In vitro infectivity was shown in KU812Ep6 cells at 24 h post-infection by in situ hybridisation and amplification assays for viral DNA and RNAs. Plasma pools with a viral load in the range of 6.00 x 10(3)-8.96 x 10(4) geq/ml did not show infectivity when inoculated into KU812Ep6 cells. Medium-high titres of IgG antibodies anti-B19 were detectable in all the plasma pools and the neutralising activity associated with specific IgG anti-B19 may explain the lack of infectivity of plasma pools contaminated with a low viral load. In conclusion, in situ hybridisation and amplification assays for viral DNA and RNAs in KU812Ep6 cells inoculated with plasma pools can be valid assays to test for the presence of infectious virus in the production of B19-safe material.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Samplesmentioning

confidence: 99%

Section: B19 Dna Titration By Competitive Pcr Assaymentioning

confidence: 99%

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Molecular testing for detection of in vitro infectivity of plasma pools contaminated with B19 virus

Bonvicini

Gallinella

Cricca

et al. 2004

Journal of Medical Virology

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“…In primary PVB 19 infection, the viral load may be as high as 10 12 to 10 14 DNA copies/ml in immunocompetent individuals (e.g. blood donors) [Weimer et al, 2001;Blumel et al, 2002;Gallinella et al, 2002], which may be even higher in the immunocompromised. In such patients, their chemotherapeutic immunosuppression may also be reduced, to allow the host immune system to clear the virus naturally [Lui et al, 2001;Eid et al, 2006].…”

Section: Discussionmentioning

confidence: 99%

Dose‐by‐dose virological and hematological responses to intravenous immunoglobulin in an immunocompromised patient with persistent parvovirus B19 infection

Tang

Lau

Wong

et al. 2007

Journal of Medical Virology

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A 42-year-old male with stage IV mantle cell lymphoma received chemotherapy and autologous peripheral blood stem cell transplantation. He developed pancytopaenia, and bone marrow examination indicated a parvovirus B19 (PVB 19)-induced red cell aplasia, confirmed by virological tests. Multiple doses of intravenous immunoglobulin (IVIG) were given over the following months, with blood samples being taken after each dose for quantitative PVB 19 DNA and hematological testing to assess the response. Each dose of IVIG produced a 1-3 log(10) drop in PVB 19 DNA levels. Eventually, after the fifth dose of IVIG, the PVB 19 DNA was reduced to <10 copies/ml serum, with a gradual improvement in his hematological parameters. This report demonstrates how close monitoring of the virological and hematological response to IVIG therapy for persistent PVB 19 infection in an immunocompromised patient can optimize the usage of this relatively expensive, and sometimes scarce intervention.

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A new primer set improves the efficiency of competitive PCR–ELISA for the detection of B19 DNA

Bonvicini

Gallinella

Cricca

et al. 2004

Journal of Clinical Virology

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Analysis of B19 virus contamination in plasma pools for manufacturing, by using a competitive polymerase chain reaction assay

Abstract: The competitive PCR assay developed proved to be effective for discrimination of the B19 virus contamination level in screening of plasma pools for manufacturing.

Cited by 12 publications

References 41 publications

Molecular testing for detection of in vitro infectivity of plasma pools contaminated with B19 virus

Molecular testing for detection of in vitro infectivity of plasma pools contaminated with B19 virus

Dose‐by‐dose virological and hematological responses to intravenous immunoglobulin in an immunocompromised patient with persistent parvovirus B19 infection

A new primer set improves the efficiency of competitive PCR–ELISA for the detection of B19 DNA

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