Diagnostic Virology Laboratory (DVL)
The USGS National Wildlife Health Center's Diagnostic Virology Laboratory (DVL) detects existing and identifies emerging pathogens of wildlife.
The DVL uses molecular testing including next generation sequencing (NGS) for rapid test turn around and is particularly skilled in virus isolation of novel pathogens. The laboratory is part of a nation-wide passive wildlife mortality surveillance program and regularly participates in large-scale active surveillance for specific diseases. This laboratory detected the introductions of West Nile Virus, highly pathogenic avian influenza virus, and spring viremia of carp virus into North America. The laboratory is currently testing for SARS-CoV-2, which causes COVID-19 in people, and rabbit hemorrhagic disease virus 2 in suitable animals. The DVL participates in the Federal Select Agent Program and is a member of the National Animal Health Laboratory Network and conducts research on viral phylogenetics, pathology, and disease ecology.
Virology Laboratory Capabilities
Facility Capabilities
- Biological Safety Level 3 (BSL-3) laboratory with dedicated isolation rooms
- Restricted access for Select Agents
- Dedicated extraction, molecular assay set up, and results analysis areas
- High-capacity sequence analysis workstation and access to high performance computing clusters
- Extensive archive of wildlife tissues and virus isolates
Molecular Diagnostic Capabilities
- Multiple manual and robotic nucleic extraction platforms
- Conventional and real time thermal cyclers for PCR and RT-PCR tests
- Access to overnight Sanger sequencing service
- Third generation next generation sequencing platforms
- Agent-specific diagnostic tests (see table) and broad-spectrum survey tests
Virus Isolation Capabilities
- Amphibian, reptilian, avian, and mammalian tissue culture systems available
- Commonly used cell lines include Zebrafish (ZF4), Muscovy duck embryo fibroblast (MSDEF), African green monkey kidney (Vero) plus a library of specialty cell lines appropriate to different host species
- Embryonated eggs are used to isolate avian influenza and Newcastle Disease viruses as well as avian poxviruses
- Isolated viruses are identified by morphological (electron microscope), antigenic, and molecular characteristics
- Isolated viruses are used for detailed genomic characterization as well as in experimental animal trials to characterize disease pathology, virus transmission, and host range determination
Serological Capabilities
- Crane Herpesvirus (Inclusion Body Disease of Cranes-IBDC)
- West Nile Virus (WNV), St Louis Encephalitis (SLE) Virus, and Japanese Encephalitis Virus
- Wellfleet Bay Virus (Common eider orthomyxovirus)
- Plaque assay for determination of viral titer
- Virus neutralization and microneutralization assays
Virology Diagnostic Tests
The following is a reference guide to commonly requested tests. It is not a full list of testing capabilities of the laboratories. Determination of which test will be run on any given submission will be based on case history, gross findings at necropsy, and the scientific judgment of the case pathologist.
| Test | Test Method |
|---|---|
| Adenovirus | PCR |
| African Swine Fever Virus (ASFV) | qRT-PCR |
| Aleutian Mink Disease | PCR |
| Alphaviruses including Togaviruses | RT-PCR |
| Avian Influenza Virus (AIV) | qRT-PCR, isolation in allantoic sac of embryonating eggs |
| Avian Avulavirus (AAvV, multiple serotypes, previously Avian Paramyxovirus, APMV) | RT-PCR, isolation in allantoic sac of embryonating eggs |
| Bunyavirus | RT-PCR |
| Calicivirus | RT-PCR |
| Canine Distemper Virus (CDV) | ELISA |
| Classical Swine Fever Virus (CSFV) | qRT-PCR |
| Coronavirus | RT-PCR |
| Duck Viral Enteritis (DVE) | PCR, isolation in MSDEF |
| Eastern Equine Encephalitis Virus (EEEV) | RT-PCR, isolation in VERO |
| Flavivirus | RT-PCR, isolation in VERO |
| Foot and Mouth Disease Virus (FMDV) | RT-PCR |
| Herpesvirus | PCR, isolation in MSDEF |
| Highland J Virus | RT-PCR, isolation in VERO |
| Inclusion Body Disease of Crane Virus (IBDCV) | Serum neutralization |
| Morbillivirus | RT-PCR |
| Newcastle Disease Virus (NDV, AAvV-1) | qRT-PCR, isolation in allantoic sac of embryonating eggs |
| Parvovirus, including CPV | PCR, CPV ELISA |
| Poecivirus associated with beak deformation | RT-PCR |
| Poxvirus | PCR, isolation in chorioallantoic membrane, MSDEF |
| Rabies | Sent out |
| Rabbit Hemorrhagic Disease Virus 2 (RHDV2) | rRT-PCR |
| Ranavirus, including FV3 | qPCR, isolation in ZF4 |
| Rhabdovirus | RT-PCR |
| Reovirus | PCR, isolate in VERO |
| Reticuloendotheliosis Virus (REV) | PCR, Isolate in VERO |
| SARS-CoV-2 | qRT-PCR, isolate in VERO |
| Turtle Fraservirus 1 (TFV1) | PCR |
| Viral Hemorrhagic Septicemia Virus (VHSV) | RT-PCR, isolate in VERO |
| West Nile Virus (WNV) | qRT-PCR, PRNT |
