Development and evaluation of ELISA procedures to detect antibodies against the major envelope protein (GL) of equine arteritis virus

Nugent, J.; Sinclair, R.; DeVries, Antoine A.F.; Eberhardt, Ruth Y; Castillo‐Olivares, Javier; Davis-Poynter, Nicholas; Rottier, Peter J. M.; Mumford, J. A.

doi:10.1016/s0166-0934(00)00231-7

Cited by 33 publications

(30 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Castillo-Olivares et al [17] developed a candidate live marker vaccine (EAV-G L ∆) for EAV by deletion of the major virus neutralising domain of the G L protein. A peptide, contained within the deleted sequence forms the basis of a discriminating ELISA assay [71]. The vaccine was well tolerated by ponies, although the vaccine virus could be easily isolated from swabs and leucocytes after intra-nasal inoculation.…”

Section: Novel Vaccination Strategies For Equine Viral Arteritismentioning

confidence: 99%

Equine viral vaccines: the past, present and future

2004

View full text Add to dashboard Cite

-The increasing international movement of horses combined with the relaxation of veterinary regulations has resulted in an increased incidence of equine infectious diseases. Vaccination, along with management measures, has become the primary method for the effective control of these diseases. Traditionally modified live and inactivated vaccines have been used and these vaccines have proven to be very successful in preventing disease. However, there are a number of equine infectious diseases for which conventional technology has shown its limitations. The advent of recombinant technology has stimulated the development of second generation vaccines, including gene deleted mutants, live vectored vaccines and DNA vaccines. These vaccines have in common that protective antigens are endogenously processed and presented along the molecules of the MHC I and MHC II complex, resulting in the stimulation of both humoral and cell-mediated immune responses similar to natural infection. The present paper provides a review of the vaccines being employed today against the most important equine viral diseases followed by a summary of new developments that are expected to bring improved vaccines to the market in the foreseeable future.

show abstract

Section: Novel Vaccination Strategies For Equine Viral Arteritismentioning

confidence: 99%

Equine viral vaccines: the past, present and future

2004

View full text Add to dashboard Cite

show abstract

“…11,18,19 Although natural outbreaks represent true dynamic situations, the timing of initial sample collection to determine serologic responses can vary depending on the identification of the initial index case(s). The present study showed that 26% of study horses had detectable EqCoV antibodies at the time the acute serum samples were collected, with group seroprevalences varying depending on the disease and qPCR status.…”

Section: Equid Herpesvirus 1 Equid Herpesvirus 4 Equine Influenza Amentioning

confidence: 99%

Development of an equine coronavirus–specific enzyme-linked immunosorbent assay to determine serologic responses in naturally infected horses

2016

View full text Add to dashboard Cite

Equine coronavirus (EqCoV) infection has been documented in most reports through quantitative qPCR analysis of feces and viral genome sequencing. Although qPCR is used to detect antigen during the acute disease phase, there is no equine-specific antibody test available to study EqCoV seroprevalence in various horse populations. We developed an enzyme-linked immunosorbent assay (ELISA) targeting antibodies to the spike (S) protein of EqCoV and validated its use, using acute and convalescent sera from 83 adult horses involved in 6 outbreaks. The EqCoV S protein–based ELISA was able to reliably detect antibodies to EqCoV in naturally infected horses. The greatest seroconversion rate was observed in horses with clinical signs compatible with EqCoV infection and EqCoV qPCR detection in feces. The EqCoV S protein–based ELISA could be used effectively for seroepidemiologic studies in order to better characterize the overall infection rate of EqCoV in various horse populations.

show abstract

“…Antibody responses to two G L -derived antigens were analyzed by ELISA in accordance with the procedures described by Nugent et al (49) with minor modifications. The antigens chosen were an ovalbumin-conjugated synthetic peptide representing aa 81 to 106 of G L (G L -OVA), a region deleted in EAV-G L ⌬, and a six-histidine-tagged recombinant protein expressing the complete G L ectodomain (residues 18 to 122; G L -6His), both described earlier (49).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…The availability of G L -based ELISAs that Nugent et al established recently (49) allowed us to analyze the antibody responses directed against the entire G L ectodomain (residues 18 to 122) and against a part (residues 81 to 106) of the domain deleted in EAV-G L ⌬ (residues 66 to 112). The former test uses as an antigen a His-tagged bacterial expression product (G L -6His), while in the latter assay an ovalbumin-coupled synthetic peptide (G L -OVA) is applied to the ELISA plates.…”

Section: Immunization Of Ponies With Eav-gmentioning

confidence: 99%

“…It is also known that the entire G L ectodomain of EAV may be replaced by the corresponding domains from other arteriviruses and still retain viability (24). Furthermore, the peptide antigen G L -OVA (aa 81 to 106), which forms the basis of an already-developed diagnostic enzyme-linked immunosorbent assay (ELISA) (49), is contained within the deleted sequence, theoretically permitting the serological discrimination between vaccinated and infected animals. We further wished to test whether removal of the immunodominant region of G L would affect the capacity of the deletion mutant virus to induce VNAb and, if so, whether the resulting immune response would protect against challenge EAV infection.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Generation of a Candidate Live Marker Vaccine for Equine Arteritis Virus by Deletion of the Major Virus Neutralization Domain

Castillo‐Olivares

Wieringa

Bakonyi

et al. 2003

J Virol

Self Cite

View full text Add to dashboard Cite

Equine arteritis virus (EAV) is an enveloped plus-strand RNA virus of the family. By using an infectious cDNA we have now generated, in the controlled background of a nonvirulent virus, a mutant EAV from which this immunodominant domain was deleted. This virus, EAV-G L ⌬, replicated to normal titers in culture cells, although at a slower rate than wild-type EAV, and caused an asymptomatic infection in ponies. The antibodies induced neutralized the mutant virus efficiently in vitro but reacted poorly to wild-type EAV strains. Nevertheless, when inoculated subsequently with virulent EAV, the immunized animals, in contrast to nonvaccinated controls, were fully protected against disease; replication of the challenge virus occurred briefly at low though detectable levels. The levels of protection achieved suggest that an immune effector mechanism other than VNAb plays an important role in protection against infection. As expected, infection with EAV-G L ⌬ did not induce a measurable response in our G Lpeptide ELISA while the challenge infection of the animals clearly did. EAV-G L ⌬ or similar mutants are therefore attractive marker vaccine candidates, enabling serological discrimination between vaccinated and wild-type virus-infected animals.Equine arteritis virus (EAV), a plus-strand RNA virus of the family Arteriviridae (order Nidovirales), is a worldwide pathogen for horses and donkeys. The virus was first isolated from lung tissue of fetuses aborted during an outbreak in Ohio in 1953 (25, 27) and became the prototype arterivirus; it was joined later by the lactate dehydrogenase-elevating virus of mice, simian hemorrhagic fever virus, and porcine reproductive and respiratory syndrome virus. Though the virus infects cells from quite a variety of origins in vitro, its host range in nature is narrowly restricted to equids (17,19,36,52,54).In the horse clinical signs of infection vary widely and appear to depend on the particular strain and dose of the virus, the age and physical condition of the animal, and possibly also the route of infection (58, 60). While epidemiological studies indicate that natural infections often occur asymptomatically (16,46,61), affected animals may develop moderate-to-severe symptoms as well. Lethal infections of horses with EAV have been reported only under experimental conditions (12, 29).While infected horses usually recover after cessation of viremia, foal death as a result of rapidly progressive bronchointerstitial pneumonia and intestinal necrosis can occur in young and adolescent pregnant mares. In mares EAV infection does not seem to affect fertility. Stallions may, however, temporarily exhibit fertility problems due to a reduced production of morphologically normal sperm cells during the first months after infection (48).Transmission of EAV occurs via the respiratory route through direct contact with infectious aerosolized nasopharyngeal secretions from acutely infected horses or other secretions such as urine; feces and vaginal fluids; and fetal, placental, and amniotic materia...

show abstract

Development and evaluation of ELISA procedures to detect antibodies against the major envelope protein (GL) of equine arteritis virus

Cited by 33 publications

References 43 publications

Equine viral vaccines: the past, present and future

Equine viral vaccines: the past, present and future

Development of an equine coronavirus–specific enzyme-linked immunosorbent assay to determine serologic responses in naturally infected horses

Generation of a Candidate Live Marker Vaccine for Equine Arteritis Virus by Deletion of the Major Virus Neutralization Domain

Contact Info

Product

Resources

About