A novel parvovirus was identified as a cell culture contaminant by metagenomic analysis. Droplet ... more A novel parvovirus was identified as a cell culture contaminant by metagenomic analysis. Droplet digital PCR (ddPCR) was used to determine viral loads in the cell culture supernatant and further analysis, by ddPCR and DNA sequencing, demonstrated that fetal bovine serum (FBS) used during cell culture was the source of the parvovirus contamination. The FBS contained ~ 50,000 copies of the novel parvovirus DNA per ml of serum. The viral DNA was resistant to DNAse digestion. Near-full length sequence of the novel parvovirus was determined. Phylogenetic analysis demonstrated that virus belongs to the Copiparvovirus genus, being most closely related to bovine parvovirus 2 (BPV2) with 41% identity with the non-structural protein NS1 and 47% identity with the virus capsid protein of BPV2. A screen of individual and pooled bovine sera identified a closely related variant of the novel virus in a second serum pool. For classification purposes, the novel virus has been designated bovine copipa...
Background: Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related members of the ... more Background: Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related members of the Semliki Forest complex within the genus alphavirus and are transmitted by arthropods, causing acute febrile illness in humans. CHIKV has spread to almost all continents, whereas autochthonous MAYV infections have been reported in South America and in the Caribbean. Nevertheless, there was concern about potential spread of MAYV to other regions similar to CHIKV in the past. The risk for transmission of emerging viruses by blood transfusion and the safety of plasma-derived medicinal products (PDMPs) are constant concerns. The manufacturing processes of PDMPs include procedures to inactivate/remove viruses. Methods: In this study, we investigated the reduction of MAYV and CHIKV by heat inactivation in various matrices, solvent/detergent treatment and nanofiltration. Results: Unexpectedly, MAYV was significantly more resistant to heat and solvent/detergent treatment compared to CHIKV. Howeve...
PDA journal of pharmaceutical science and technology
This report provides a high-level summary of the key outcomes and gaps based on the research pres... more This report provides a high-level summary of the key outcomes and gaps based on the research presented at the Viral Clearance Symposium 2017 and identifies new areas for future study and improvements.The 2017 conference structure extended the framework from the preceding conferences, focusing on the key gaps and associated developments and including the additional focus areas of facility risk mitigation and continuous processing, and ways to improve the efficiency of the overall adventitious agent strategy. This report provides a high-level summary of the key outcomes and gaps based on the research presented at the Viral Clearance Symposium 2017 and identifies new areas for future study and improvements.
We report here the genome sequence of a hepatitis E virus (HEV) strain from a chronically infecte... more We report here the genome sequence of a hepatitis E virus (HEV) strain from a chronically infected immunodeficient patient. Full-length sequence analysis revealed a distinct HEV strain, of a tentative new subgenotype, clustering with viruses from rabbits. It is a World Health Organization reference strain for validation of nucleic acid testing.
We conducted an external quality assessment of Zika virus molecular diagnostic tests in Brazil us... more We conducted an external quality assessment of Zika virus molecular diagnostic tests in Brazil using a new Zika virus standard. Of 15 laboratories, 73% showed limited sensitivity and specificity. Viral load estimates varied significantly. Continuous quality assurance is needed to adequately estimate risk for Zika virus-associated disease and determine patient care.
We report here the sequence of a genotype 2a reference strain of hepatitis E virus (HEV), develop... more We report here the sequence of a genotype 2a reference strain of hepatitis E virus (HEV), developed on behalf of the World Health Organization. The HEV reference strain is intended for use in assays based on nucleic acid amplification for the validation of HEV RNA detection.
Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin, Jan 20, 2017
The public health implications of hepatitis E virus (HEV) in Europe have changed due to increasin... more The public health implications of hepatitis E virus (HEV) in Europe have changed due to increasing numbers of hepatitis E cases and recent reports of chronic, persistent HEV infections associated with progression to cirrhosis in immunosuppressed patients. The main infectious risk for such immunosuppressed patients is exposure to undercooked infected pork products and blood transfusion. We summarised the epidemiology of HEV infections among blood donors and also outlined any strategies to prevent transfusion-transmitted HEV, in 11 European countries. In response to the threat posed by HEV and related public and political concerns, most of the observed countries determined seroprevalence of HEV in donors and presence of HEV RNA in blood donations. France, Germany, Spain and the United Kingdom (UK) reported cases of transfusion-transmitted HEV. Ireland and the UK have already implemented HEV RNA screening of blood donations; the Netherlands will start in 2017. Germany and France perfor...
It is now recognized that blood donors may be silently infected with hepatitis E virus (HEV) and ... more It is now recognized that blood donors may be silently infected with hepatitis E virus (HEV) and that plasma pools used in the manufacture of plasma-derived medicinal products may also contain detectable virus RNA. The occurrence of HEV-infected blood and plasma donors can vary considerably depending on local epidemiology. Manufacturing plasma pools from North America, Europe, the Middle East, and Asia were examined for the presence of HEV using transcription-mediated amplification of HEV RNA; confirmatory testing was performed using real-time reverse transcription polymerase chain reaction and sequencing. A total of 484 pools were tested. Asian pools were most frequently positive for HEV RNA and had higher viral loads, although none exceeding 300 IU/mL, and the sequenced strains (n = 5) clustered with Genotype 4, including one significantly divergent sequence. Only HEV Genotype 3 was identified in North American (n = 5) and European (n = 5) pools. There was no evidence of HEV in an...
We report here the sequence of a candidate reference strain of Zika virus (ZIKV) developed on beh... more We report here the sequence of a candidate reference strain of Zika virus (ZIKV) developed on behalf of the World Health Organization (WHO). The ZIKV reference strain is intended for use in nucleic acid amplification (NAT)-based assays for the detection and quantification of ZIKV RNA.
Zika virus (ZIKV) is an emerging mosquito-borne Flavivirus of major public health concern. The po... more Zika virus (ZIKV) is an emerging mosquito-borne Flavivirus of major public health concern. The potential for ZIKV transmission by blood transfusion has been demonstrated; however, inactivation or removal of ZIKV during the manufacture of plasma-derived medicinal products has not been specifically investigated. Inactivation of ZIKV by pasteurization and solvent/detergent (S/D) treatment was investigated by spiking high-titer ZIKV stocks into human serum albumin and applying either heat or adding different mixtures of S/D reagents and assaying for infectious virus particles. Removal of ZIKV was evaluated using filters of differing pore sizes (75, 40, 35, and 19 nm), assaying for infectious virus and RNA. Electron microscopy was performed to determine the size of ZIKV particles. Neutralization of virus infectivity by immunoglobulins was investigated. ZIKV was effectively and rapidly inactivated by liquid heat treatment as well as by various mixtures of S/D reagents with reduction facto...
A novel parvovirus was identified as a cell culture contaminant by metagenomic analysis. Droplet ... more A novel parvovirus was identified as a cell culture contaminant by metagenomic analysis. Droplet digital PCR (ddPCR) was used to determine viral loads in the cell culture supernatant and further analysis, by ddPCR and DNA sequencing, demonstrated that fetal bovine serum (FBS) used during cell culture was the source of the parvovirus contamination. The FBS contained ~ 50,000 copies of the novel parvovirus DNA per ml of serum. The viral DNA was resistant to DNAse digestion. Near-full length sequence of the novel parvovirus was determined. Phylogenetic analysis demonstrated that virus belongs to the Copiparvovirus genus, being most closely related to bovine parvovirus 2 (BPV2) with 41% identity with the non-structural protein NS1 and 47% identity with the virus capsid protein of BPV2. A screen of individual and pooled bovine sera identified a closely related variant of the novel virus in a second serum pool. For classification purposes, the novel virus has been designated bovine copipa...
Background: Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related members of the ... more Background: Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related members of the Semliki Forest complex within the genus alphavirus and are transmitted by arthropods, causing acute febrile illness in humans. CHIKV has spread to almost all continents, whereas autochthonous MAYV infections have been reported in South America and in the Caribbean. Nevertheless, there was concern about potential spread of MAYV to other regions similar to CHIKV in the past. The risk for transmission of emerging viruses by blood transfusion and the safety of plasma-derived medicinal products (PDMPs) are constant concerns. The manufacturing processes of PDMPs include procedures to inactivate/remove viruses. Methods: In this study, we investigated the reduction of MAYV and CHIKV by heat inactivation in various matrices, solvent/detergent treatment and nanofiltration. Results: Unexpectedly, MAYV was significantly more resistant to heat and solvent/detergent treatment compared to CHIKV. Howeve...
PDA journal of pharmaceutical science and technology
This report provides a high-level summary of the key outcomes and gaps based on the research pres... more This report provides a high-level summary of the key outcomes and gaps based on the research presented at the Viral Clearance Symposium 2017 and identifies new areas for future study and improvements.The 2017 conference structure extended the framework from the preceding conferences, focusing on the key gaps and associated developments and including the additional focus areas of facility risk mitigation and continuous processing, and ways to improve the efficiency of the overall adventitious agent strategy. This report provides a high-level summary of the key outcomes and gaps based on the research presented at the Viral Clearance Symposium 2017 and identifies new areas for future study and improvements.
We report here the genome sequence of a hepatitis E virus (HEV) strain from a chronically infecte... more We report here the genome sequence of a hepatitis E virus (HEV) strain from a chronically infected immunodeficient patient. Full-length sequence analysis revealed a distinct HEV strain, of a tentative new subgenotype, clustering with viruses from rabbits. It is a World Health Organization reference strain for validation of nucleic acid testing.
We conducted an external quality assessment of Zika virus molecular diagnostic tests in Brazil us... more We conducted an external quality assessment of Zika virus molecular diagnostic tests in Brazil using a new Zika virus standard. Of 15 laboratories, 73% showed limited sensitivity and specificity. Viral load estimates varied significantly. Continuous quality assurance is needed to adequately estimate risk for Zika virus-associated disease and determine patient care.
We report here the sequence of a genotype 2a reference strain of hepatitis E virus (HEV), develop... more We report here the sequence of a genotype 2a reference strain of hepatitis E virus (HEV), developed on behalf of the World Health Organization. The HEV reference strain is intended for use in assays based on nucleic acid amplification for the validation of HEV RNA detection.
Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin, Jan 20, 2017
The public health implications of hepatitis E virus (HEV) in Europe have changed due to increasin... more The public health implications of hepatitis E virus (HEV) in Europe have changed due to increasing numbers of hepatitis E cases and recent reports of chronic, persistent HEV infections associated with progression to cirrhosis in immunosuppressed patients. The main infectious risk for such immunosuppressed patients is exposure to undercooked infected pork products and blood transfusion. We summarised the epidemiology of HEV infections among blood donors and also outlined any strategies to prevent transfusion-transmitted HEV, in 11 European countries. In response to the threat posed by HEV and related public and political concerns, most of the observed countries determined seroprevalence of HEV in donors and presence of HEV RNA in blood donations. France, Germany, Spain and the United Kingdom (UK) reported cases of transfusion-transmitted HEV. Ireland and the UK have already implemented HEV RNA screening of blood donations; the Netherlands will start in 2017. Germany and France perfor...
It is now recognized that blood donors may be silently infected with hepatitis E virus (HEV) and ... more It is now recognized that blood donors may be silently infected with hepatitis E virus (HEV) and that plasma pools used in the manufacture of plasma-derived medicinal products may also contain detectable virus RNA. The occurrence of HEV-infected blood and plasma donors can vary considerably depending on local epidemiology. Manufacturing plasma pools from North America, Europe, the Middle East, and Asia were examined for the presence of HEV using transcription-mediated amplification of HEV RNA; confirmatory testing was performed using real-time reverse transcription polymerase chain reaction and sequencing. A total of 484 pools were tested. Asian pools were most frequently positive for HEV RNA and had higher viral loads, although none exceeding 300 IU/mL, and the sequenced strains (n = 5) clustered with Genotype 4, including one significantly divergent sequence. Only HEV Genotype 3 was identified in North American (n = 5) and European (n = 5) pools. There was no evidence of HEV in an...
We report here the sequence of a candidate reference strain of Zika virus (ZIKV) developed on beh... more We report here the sequence of a candidate reference strain of Zika virus (ZIKV) developed on behalf of the World Health Organization (WHO). The ZIKV reference strain is intended for use in nucleic acid amplification (NAT)-based assays for the detection and quantification of ZIKV RNA.
Zika virus (ZIKV) is an emerging mosquito-borne Flavivirus of major public health concern. The po... more Zika virus (ZIKV) is an emerging mosquito-borne Flavivirus of major public health concern. The potential for ZIKV transmission by blood transfusion has been demonstrated; however, inactivation or removal of ZIKV during the manufacture of plasma-derived medicinal products has not been specifically investigated. Inactivation of ZIKV by pasteurization and solvent/detergent (S/D) treatment was investigated by spiking high-titer ZIKV stocks into human serum albumin and applying either heat or adding different mixtures of S/D reagents and assaying for infectious virus particles. Removal of ZIKV was evaluated using filters of differing pore sizes (75, 40, 35, and 19 nm), assaying for infectious virus and RNA. Electron microscopy was performed to determine the size of ZIKV particles. Neutralization of virus infectivity by immunoglobulins was investigated. ZIKV was effectively and rapidly inactivated by liquid heat treatment as well as by various mixtures of S/D reagents with reduction facto...
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