Recommendations for the introduction of metagenomic high-throughput sequencing in clinical virology, part I:Wet lab procedure

Metagenomic high-throughput sequencing (mHTS) is a hypothesis-free, universal pathogen detection technique for determination of the DNA/RNA sequences in a variety of sample types and infectious syndromes. mHTS is still in its early stages of translating into clinical application. To support the development, implementation and standardization of mHTS procedures for virus diagnostics, the European Society for Clinical Virology (ESCV) Network on Next-Generation Sequencing (ENNGS) has been established. The aim of ENNGS is to bring together professionals involved in mHTS for viral diagnostics to share methodologies and experiences, and to develop application recommendations. This manuscript aims to provide practical recommendations for the wet lab procedures necessary for implementation of mHTS for virus diagnostics and to give recommendations for development and validation of laboratory methods, including mHTS quality assurance, control and quality assessment protocols

Recommendations for the introduction of metagenomic high-throughput sequencing in clinical virology, part I: Wet lab procedure

Metagenomic high-throughput sequencing (mHTS) is a hypothesis-free, universal pathogen detection technique for determination of the DNA/RNA sequences in a variety of sample types and infectious syndromes. mHTS is still in its early stages of translating into clinical application. To support the development, implementation and standardization of mHTS procedures for virus diagnostics, the European Society for Clinical Virology (ESCV) Network on Next-Generation Sequencing (ENNGS) has been established. The aim of ENNGS is to bring together professionals involved in mHTS for viral diagnostics to share methodologies and experiences, and to develop application recommendations. This manuscript aims to provide practical recommendations for the wet lab procedures necessary for implementation of mHTS for virus diagnostics and to give recommendations for development and validation of laboratory methods, including mHTS quality assurance, control and quality assessment protocols

Recommendations for the introduction of metagenomic high-throughput sequencing in clinical virology, part I: Wet lab procedure

Metagenomic high-throughput sequencing (mHTS) is a hypothesis-free, universal pathogen detection technique for determination of the DNA/RNA sequences in a variety of sample types and infectious syndromes. mHTS is still in its early stages of translating into clinical application. To support the development, implementation and standardization of mHTS procedures for virus diagnostics, the European Society for Clinical Virology (ESCV) Network on Next-Generation Sequencing (ENNGS) has been established. The aim of ENNGS is to bring together professionals involved in mHTS for viral diagnostics to share methodologies and experiences, and to develop application recommendations. This manuscript aims to provide practical recommendations for the wet lab procedures necessary for i

Serology Identifies LIPyV as a Feline Rather than a Human Polyomavirus.

The number of identified human polyomaviruses (HPyVs) has increased steadily over the last decade. Some of the novel HPyVs have been shown to cause disease in immunocompromised individuals. The Lyon-IARC polyomavirus (LIPyV) belonging to species Alphapolyomavirus quardecihominis was identified in 2017 in skin and saliva samples from healthy individuals. Since its initial discovery, LIPyV has rarely been detected in human clinical samples but has been detected in faeces from cats with diarrhoea. Serological studies show low LIPyV seroprevalence in human populations. To investigate the possibility that LIPyV is a feline rather than a human polyomavirus, we compared serum IgG responses against the VP1 major capsid protein of LIPyV and 13 other HPyVs among cats (n = 40), dogs (n = 38) and humans (n = 87) using an in-house immunoassay. Seropositivity among cats was very high (92.5%) compared to dogs (31.6%) and humans (2.3%). Furthermore, the median antibody titres against LIPyV were 100-10,000x higher in cats compared to dogs and humans. In conclusion, the high prevalence and intensity of measured seroresponses suggest LIPyV to be a feline rather than a human polyomavirus. Whether LIPyV infection induces diarrhoea or other symptoms in cats remains to be established

Seroprevalence of fourteen human polyomaviruses determined in blood donors.

The polyomavirus family currently includes thirteen human polyomavirus (HPyV) species. In immunocompromised and elderly persons HPyVs are known to cause disease, such as progressive multifocal leukoencephalopathy (JCPyV), haemorrhagic cystitis and nephropathy (BKPyV), Merkel cell carcinoma (MCPyV), and trichodysplasia spinulosa (TSPyV). Some recently discovered polyomaviruses are of still unknown prevalence and pathogenic potential. Because HPyVs infections persist and might be transferred by blood components to immunocompromised patients, we studied the seroprevalence of fourteen polyomaviruses in adult Dutch blood donors. For most polyomaviruses the observed seroprevalence was high (60-100%), sometimes slightly increasing or decreasing with age. Seroreactivity increased with age for JCPyV, HPyV6 and HPyV7 and decreased for BKPyV and TSPyV. The most recently identified polyomaviruses HPyV12, NJPyV and LIPyV showed low overall seroprevalence (~5%) and low seroreactivity, questioning their human tropism. Altogether, HPyV infections are common in Dutch blood donors, with an average of nine polyomaviruses per subject

Development and evaluation of a broad bead-based multiplex immunoassay to measure IgG seroreactivity against human polyomaviruses

International audienceThe family of polyomaviruses, which cause severe disease in immunocompromised hosts, has expanded substantially in recent years. To accommodate measurement of IgG seroresponses against all currently known human polyomaviruses (HPyVs), including the Lyon IARC polyomavirus (LIPyV), we extended our custom multiplex beadbased HPyV immunoassay and evaluated the performance of this pan-HPyV immunoassay. The VP1 proteins of 15 HPyVs belonging to 13 Polyomavirus species were expressed as recombinant glutathione S-transferase (GST) fusion proteins and coupled to fluorescent Luminex beads. Sera from healthy blood donors and immunocompromised kidney transplant recipients were used to analyze seroreactivity against the different HPyVs. For BK polyomavirus (BKPyV), the GST-VP1 fusion protein-directed seroresponses were compared to those obtained against BKPyV VP1 virus-like particles (VLP). Seroreactivity against most HPyVs was common and generally high in both test populations. Low seroreactivity against HPyV9, HPyV12, New Jersey PyV, and LIPyV was observed. The assay was reproducible (Pearson’s r2 > 0.84, P ρ = 0.823, P < 0.001). The bead-based pan-HPyV multiplex immunoassay is a reliable tool to determine HPyVspecific seroresponses with high reproducibility and specificity and is suitable for use in seroepidemiological studies

JC and Human polyomavirus 9 after kidney transplantation: An exploratory serological cohort study

Introduction: Human polyomaviruses (HPyVs) cause disease in immunocompromised patients. BK polyomavirus (BKPyV) for instance persistently infects the kidneys. In kidney transplant recipients, (KTRs) BKPyV can cause allograft nephropathy. JCPyV, MCPyV, TSPyV and HPyV9 reside in the kidneys too, or have been detected in urine. In this study, we investigate exposure to JCPyV, MCPyV, TSPyV and HPyV9 after kidney transplantation by serological means. Materials and methods: Serum samples from 310 KTR collected before and 6 months after transplantation (n = 620), from 279 corresponding kidney donors collected before transplantation, and from blood donor controls collected one year apart (n = 174) were assessed for HPyV species-specific IgG responses using a multiplex immunoassay. KTR HPyV IgG kinetics were compared to those of healthy blood donors by linear mixed modeling, and related to those of their donors by linear regression. Results: In the KTR, increased IgG levels during follow-up were observed for JCPyV (14.8%), MCPyV (7.1%), TSPyV (10.6%), and for HPyV9 (8.1%), while blood donor antibody levels remained stable. Seroconversion was observed for JCPyV (6.5%), MCPyV (2.3%), TSPyV (1.3%), and for HPyV9 (6.5%). The linear mixed model analysis showed that antibody increase was significant for JCPyV (p < 0.001) and HPyV9 (p < 0.001). Post-transplant JCPyV and HPyV9 antibody responses were associated with donor antibody levels against these HPyVs, respectively. Conclusions: KTR are exposed to JCPyV and HPyV9 after transplantation. Whether the allograft serves as the source, as indicated by the donor serostatus association, deserves further study

Los cinco millones de huevos de aves en las colecciones de museos del mundo son un recurso invaluable y subutilizado

The ~1.97 million egg sets (~5 million eggs) housed in museums have not been used in proportion to their availability. We highlight the wide variety of scientific disciplines that have used egg collections and the geographic locations and sizes of these collections, to increase awareness of the importance of egg collections, improve their visibility to the scientific community, and suggest that they offer a wealth of data covering large spatial scales and long time series for broad investigations into avian biology. We provide a brief history of egg collections and an updated list of museums/ institutions with egg collections worldwide. We also review the limitations, challenges, and management of egg collections, and summarize recent literature based on historical and recent museum egg materials.Los casi 1.97 millones de conjuntos de huevos (~5 millones de huevos) depositados en colecciones no han sido utilizados en proporción a su abundancia. En este trabajo, destacamos la subutilización de las colecciones de huevos a pesar de generar datos en amplias escalas espaciales y temporales, para incrementar la conciencia sobre la importancia de las colecciones de huevos, mejorar su visibilidad para la comunidad científica, y mostrar que las colecciones de huevos ofrecen una abundancia de datos a gran escala espacial y temporal para la investigación de la biología de las aves. Proporcionamos una breve historia de las colecciones de huevos y una lista actualizada de museos/instituciones con colecciones de huevos en todo el mundo. También discutimos las limitaciones, los desafíos y el manejo de este tipo de colecciones, y fornecemos un resumen da literatura que utiliza dados históricos y recientes de colecciones de huevos.Fil: Marini, Miguell Ângelo. Universidade do Brasília; BrasilFil: Hall, Linnea. Western Foundation Of Vertebrate Zoology; Estados UnidosFil: Bates, John. Field Museum Of Natural History; Estados UnidosFil: Steinheimer, Frank. Martin Luther University Halle-wittenberg; AlemaniaFil: McGowan, Robert. National Museums Scotland; Reino UnidoFil: Silveira, Luis Fabio. Universidade de Sao Paulo; BrasilFil: Lijtmaer, Dario Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Tubaro, Pablo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Córdoba Córdoba, Sergio. Instituto de Investigación de Recursos Biológicos Alexander von Humboldt; ColombiaFil: Gamauf, Anita. Naturhistorisches Museum; AustriaFil: Greeney, Harold F.. Yanayacu Biological Station; EcuadorFil: Schweizer, Manuel. Naturhistorisches Museum; SuizaFil: Kamminga, Pepijn. Naturalis Biodiversity Center; Países BajosFil: Cibois, Alice. Muséum d’histoire naturelle de la Ville de Genève; SuizaFil: Vallotton, Laurent. Muséum d’histoire naturelle de la Ville de Genève; SuizaFil: Russell, Douglas. Natural History Museum; Reino UnidoFil: Robinson, Scott K.. University Of Florida. Florida Museum Of History; Estados UnidosFil: Sweet, Paul R.. American Museum of Natural History; Estados UnidosFil: Frahnert, Sylke. Staatliches Museum fur Naturkunde Stuttgart; AlemaniaFil: Corado, René. Western Foundation of Vertebrate Zoology; Estados UnidosFil: Heming, Neander Marcel. Universidade Estadual de Santa Cruz; Brasil. Universidade do Brasília; Brasi

The five million bird eggs in the world’s museum collections are an invaluable and underused resource

The ~1.97 million egg sets (~5 million eggs) housed in museums have not been used in proportion to their availability. We highlight the wide variety of scientific disciplines that have used egg collections and the geographic locations and sizes of these collections, to increase awareness of the importance of egg collections, improve their visibility to the scientific community, and suggest that they offer a wealth of data covering large spatial scales and long time series for broad investigations into avian biology. We provide a brief history of egg collections and an updated list of museums/institutions with egg collections worldwide. We also review the limitations, challenges, and management of egg collections, and summarize recent literature based on historical and recent museum egg materials