Qualitative evaluation of selective tests for ...

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Title Qualitative evaluation of selective tests for detection of Neospora hughesi antibodies in serum and cerebrospinal fluid of experimentally infected horses
Author(s) A. E. Packham, P. A. Conrad, W. D. Wilson, L. V. Jeanes, K. W. Sverlow, I. A. Gardner, B. M. Daft, A. E. Marsh, B. L. Blagburn, G. L. Ferraro, B. C. Barr
Journal Journal of Parasitology
Date 2002
Volume 88
Issue 6
Start page 1239
End page 1246
Abstract Neospora hughesi is a newly recognized protozoan pathogen in horses that causes a myeloencephalitis similar to Sarcocystis neurona. There are no validated serological tests using the gold standard sera that are currently available to detect specific N. hughesi antibodies and, thus, no tests available to detect antemortem exposure or estimate seroprevalence in the horse. The objectives of the present study were to establish a bank of gold standard equine sera through experimental infections with N. hughesi and to assess several serological tests for the detection of related protozoan antibodies. Seven horses were inoculated with N. hughesi tachyzoites, and 7 horses received uninfected cell culture material. The horses were monitored, and blood and cerebrospinal fluid were collected repeatedly over a 4-month period. With the sera, 4 different serological techniques were evaluated, including a whole-parasite lysate enzyme-linked immunosorbent assay (ELISA), a recombinant protein ELISA, a modified direct agglutination test, and an indirect fluorescent antibody test. Qualitative and quantitative evaluation of the results showed that the N. hughesi indirect fluorescent antibody test (IFAT) consistently discriminated between experimentally infected and noninfected horses, using a cutoff of 1:640. Sera from 3 naturally infected horses had titers >1:640. Cerebrospinal fluid in all but one infected horse had very low N. hughesi IFAT titres (<1:160), starting at postinoculation day 30.
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