equine
assay data sheet
Equine Infectious Anemia
Test codes:
S0064
- Ultrasensitive qualitative detection of equine infectious
anemia proviral DNA by real time polymerase chain reaction
Equine infectious anemia (EIA), also known
as Swamp Fever, is a viral disease known to occur in all members
of equidae, including horses, mules and donkeys. Equine infectious
anemia virus (EIAV) is a member of the genus Lentivirus of
the family Retroviridae. EIAV is unique among the lentiviruses
in that the initial acute febrile response and associated
viremia are followed by recurrent cycles of the disease and,
finally, a prolonged asymptomatic period.
Acute cases experience fever and hemorrhaging
7-30 days post infection. Very few horses with this initial
fever are detected by owners. Chronic infection is marked
by episodes of fever, weight loss, depression, progressive
weakness, anemia and edema. These signs occur every two weeks
in recurring cycles.
Other symptoms that may occur during the course
of the disease include loss of appetite, frequent urination,
diarrhea, weakness, paralysis of the hindquarters, paleness
of the mucous membranes, yellowish discoloration of the conjunctiva,
small pinpoint hemorrhages beneath the tongue, rapid breathing
and accelerated pulse. Pregnant mares may abort.
The onset of these signs is often associated
with stresses such as hard work, hot weather, racing, pregnancy
or use of steroid drugs. Some animals show no clinical signs
associated with the infection and may go undetected. These
apparently healthy carriers harboring the virus are a constant
reservoir of infection.
There is currently no effective vaccine to
prevent EIAV infection. Routine serological testing and removal
of seropositive reactors limits the spread of this disease.
However, the utility of serological tests depends on the length
of time it takes for an animal to mount an immune response
and on the test’s sensitivity and sensitivity of antibody
detection. Currently, the agar gel immunodiffusion assay (the
Coggins test) is widely accepted and used for serodetection
of EIAV group specific antigen p26. Competitive enzyme-linked
immunosorbent assay (c-ELISA) and synthetic antigen ELISA
(SA-ELISA) have also been used for the serological detection
of EIA specific antibodies. A fluorescent polarization-based
diagnostic assay for detection of EIAV antibodies has also
been reported ( Tencza et al., 2000). Besides serological
assays, virus isolation can be used but it is not practical
because it is time consuming, laborious and requires specialized
technical skills to maintain primary horse macrophage cultures
for the replication of pathogenic strains of EIAV. The high
cost and slowness of virus isolation make it unsuitable for
routine monitoring.
Detection of EIAV proviral DNA in blood cells
by PCR has been shown to be more sensitive and specific than
serological assays not only in identifying subclinical EIAV-infected
horses, but also recently infected horses still in the process
of mounting an immune response (Spyrou, et al., 2003).
Utilities:
- Confirm the disease causing agent
- Shorten the time required to confirm a
clinical diagnosis of EIA virus infection.
- Ensure that horse populations are free
of EIA virus
- Early prevention of spread of the virus
- Minimize personnel exposure to the virus
- Safety monitoring of biological products
that derive from horses
References:
Spyrou, V., Papanastassopoulou, M., Psychas, V., Billinis,
C.H, Koumbati, M., Vlemmas, J. and Koptopoulos, G. (2003).
Equine infectious anemia in mules: virus isolation and pathogenicity
studies. Vet Microbiol. 95:49-59.
Tencza, S.B., Islam, K.R., Kalia, V., Nasir, M.S., Jolley,
M.E. and Montelaro, R.C. (2000). Development of a fluorescence
polarization-based diagnostic assay for equine infectious
anemia virus. J. Clin. Microbiol. 38:1854–1859.
Specimen requirements: 1
ml whole blood in EDTA (purple top) or ACD (yellow top) tube,
or 1 ml tissue, shipped overnight at room temperature; or
1 ml frozen tissue, shipped frozen.
For specimen types other than those listed
here, please call to confirm specimen acceptability and shipping
instructions.
For all specimen types, if there will be a
delay in shipping, or during very warm weather, refrigerate
specimens until shipped and ship with a cold pack unless more
stringent shipping requirements are specified. Frozen specimens
should be shipped so as to remain frozen in transit. See shipping
instructions for more information.
Turnaround time: 2 business
days
Methodology: Qualitative
real time PCR
Normal range: Nondetected