Diagnosis of Eastern Equine Encephalomyelitis Virus Infection in Horses by Immunoglobulin M and G Capture Enzyme-Linked Immunosorbent Assay

Sahu, Sudhir P.; Alstad, Arnold D.; Pedersen, Douglas D.; Pearson, James E.

doi:10.1177/104063879400600107

Cited by 39 publications

(14 citation statements)

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“…HI and VN are standard serologic assays used for diagnosis of EEE in suspect animals and humans. 2,7,9 However, interpretation of results on a single serum sample is often difficult without a supporting clinical history or if an animal has been vaccinated. 9 Detection of IgM antibodies to EEE virus may help to distinguish recent infection from previous vaccination 9 but many animals die of the disease within 24 hours of the onset of clinical signs and blood samples are not even taken.…”

Section: Discussionmentioning

confidence: 99%

“…Diagnosis of the disease generally is based on clinical history, identification of characteristic histopathologic changes in the brains of infected animals, and virus isolation. 9,10 Serologic testing also may be useful, but interpretation of hemagglutination inhibition (HI) and virus neutralization (VN) antibody titers from a single serum sample is often difficult and may be confounded by vaccination. 9 Confirmation of a histopathologic diagnosis of EEE by virus isolation is reliable, but many diagnostic laboratories are not equipped for such a procedure or are unwilling to accept the health risk to personnel.…”

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Immunohistochemical Diagnosis of Eastern Equine Encephalomyelitis

et al. 1996

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Abstract. An immunohistochemical (IHC) assay was developed for the detection of eastern equine encephalomyelitis (EEE) virus antigen in formalin-fixed, paraffin-embedded tissues. All cases of EEE diagnosed at the Michigan State University Animal Health Diagnostic Laboratory from 1991 through 1994 were evaluated. The diagnosis was based on histopathologic examination of the brain and confirmatory virus isolation. Sections of cerebrum from 26 equids and 5 birds were assessed by IHC. Histologically normal brain tissues from 2 horses and 1 pheasant and brain tissues from 2 cases of equine neurologic disease with diagnoses other than EEE served as negative controls. The IHC assay was based on standard streptavidin-biotin technology, using a commercially available kit and a monospecific polyclonal primary antibody preparation derived from murine ascitic fluid. Nineteen of 20 equids and all 5 birds positive by histopathology and virus isolation were positive for EEE virus antigen by IHC. Three equids with histologic lesions compatible with a diagnosis of EEE but negative by virus isolation also were negative for virus antigen by IHC. In 3 other equids, histopathology and IHC were positive for EEE, but virus isolation was not attempted because of contamination of the brain specimen. The IHC assay of formalin-fixed, paraffin-embedded brain tissues for EEE virus antigen is a rapid, effective test for confirming a histopathologic diagnosis of EEE, and assay results correlate well with virus isolation results.

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Immunohistochemical Diagnosis of Eastern Equine Encephalomyelitis

et al. 1996

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“…[1][2][3][4]9,11,12,14,17,20,22,24,25 Based on documented laboratory results and published reports on the immune response to EEEV, horses require 5-7 days PI to produce detectable EEEV titers. 21 Pigs and humans require at least 5 days PI and emus 7 days. 6,19,23 Extrapolating from this data, it may be postulated that dogs may require at least 5-9 days for a quantifiable amount of IgG to be detected and that the puppy with the serology antibody titer and positive ancillary tests likely harbored the virus for the period of time required for production of IgG antibodies to occur before succumbing to the disease.…”

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Eastern Equine Encephalitis in Dogs

et al. 2005

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Abstract. Eastern equine encephalitis virus (EEEV) is anAlphavirus that is endemic in the Southeastern United States. From 1993 to January 2005, the Veterinary Diagnostic and Investigational Laboratory in Tifton, Georgia, performed postmortem examinations on over 101 domestic canines exhibiting clinical neurological disturbances. In 12 of these dogs, brains were histologically suggestive of infection with EEEV. All dogs were less than 6 months of age, with no breed predilection. Clinical signs included pyrexia, depression, nystagmus, and lateral recumbency. Microscopically, brains from all 12 puppies contained infiltrates of lymphocytes, plasma cells, and histiocytes, with occasional neutrophils and random foci of astrocytosis and gliosis. There were mild to moderate perivascular infiltrates of neutrophils along with scattered lymphocytes, plasma cells, and macrophages in the meninges. Viruses isolated from brain homogenates of all 12 puppies were confirmed by indirect fluorescent antibody testing to be EEEV. Additionally, RNA extracted from the brains and viral cultures of 2 dogs were determined by a specific reverse-transcriptase polymerase chain reaction (RT-PCR) to contain EEEV. The single available serum sample exhibited a 1 : 8 serum neutralization titer to EEEV.

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“…The ELISA was performed as previously described. 34 Briefly, all wells of ELISA microplates (Immulon-1B; Dynex Technologies, Inc., Chantilly, VA) were coated with a 1:400 dilution of anti-equine IgM in freshly prepared 0.5M carbonate buffer, pH 9.6. The plates were incubated at 4°C overnight in a humidified chamber.…”

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Pathogenicity of a Venezuelan Equine Encephalomyelitis Serotype Ie Virus Isolate for Ponies

Sahu

Pedersen

Jenny

et al. 2003

The American Journal of Tropical Medicine and Hygiene

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Abstract. The enzootic or endemic strains of Venezuelan equine encephalomyelitis (VEE) virus (ID, IE, IF, and II-VI) are considered avirulent. In 1993 and 1996, outbreaks of encephalitis occurred in the horse populations in the Chiapas and Oaxaca provinces of Mexico, respectively. In both instances, enzootic VEE virus subserotype IE was isolated from brain tissues of dead horses. The present study investigated the pathogenicity of the Chiapas viral isolate (NVSL VEE IE 93-42124) in ponies. Three ponies were inoculated intradermally with 4, 5, and 6 logs, respectively, of the NVSL VEE IE 93-42124 viral isolate. All ponies showed fluctuations in body temperature, encephalitis, and other signs of infection with VEE virus. Virus was isolated only from the blood of ponies from day 1 to day 3 postinfection. Microscopic examination of hematoxylin and eosin-stained tissue sections showed mild to moderate nonsuppurative encephalitis, perivascular cuffing by mononuclear cells, gliosis, and meningoencephalitis. Antibody (IgM) to VEE virus IE was unable to differentiate between various subserotypes of VEE I viruses (serotypes IAB, IC, ID, and IF). Virus neutralizing antibody titers to heterologous VEE I viruses were 10-100-fold less than those for NVSL VEE IE 93-42124 virus and Mena II, a human isolate of VEE IE virus. The study confirmed that NVSL VEE IE 93-42124 virus, which was isolated from a brain of a horse during an outbreak of VEE in Chiapas, Mexico, was pathogenic for ponies.

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Diagnosis of Eastern Equine Encephalomyelitis Virus Infection in Horses by Immunoglobulin M and G Capture Enzyme-Linked Immunosorbent Assay

Cited by 39 publications

References 9 publications

Immunohistochemical Diagnosis of Eastern Equine Encephalomyelitis

Immunohistochemical Diagnosis of Eastern Equine Encephalomyelitis

Eastern Equine Encephalitis in Dogs

Pathogenicity of a Venezuelan Equine Encephalomyelitis Serotype Ie Virus Isolate for Ponies

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