Improvement of Western Blot Test Specificity for Detecting Equine Serum Antibodies to<i>Sarcocystis Neurona</i>

Rossano, M.G.; Mansfield, Linda S.; Kaneene, John B.; Murphy, Alice J.; Brown, Christopher M.; Schott, Harold C.; Fox, J. Carl

doi:10.1177/104063870001200105

Cited by 48 publications

(40 citation statements)

References 18 publications

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“…11 The improvement of the immunoblot analysis was based on the elimination of any cross-reactivity that may occur with Sarcocystis cruzi by preincubation of the blots in pooled bovine serum. Despite this report, the present investigation has used the original immunoblot analysis as outlined by Granstrom et al 6 The immunoblot used for this investigation is justified and is believed to provide an accurate estimate of the seroprevalences reported for the state of Oklahoma.…”

Section: Seroprevalence Of Antibodies To Sarcocystis Neurona In Equidmentioning

confidence: 99%

Seroprevalence of Antibodies toSarcocystis Neuronain Equids Residing in Oklahoma

et al. 2003

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Abstract.A sampling of equids from the state of Oklahoma produced an estimate of seroprevalence of antibody to Sarcocystis neurona to be about 89.2%. This figure represents the highest currently reported regional seroprevalence of antibody to this organism. Regional differences in seroprevalence were found in the western quadrants of the state relative to the eastern quadrants of the state, with a significantly higher seroprevalence in the eastern regions. Thoroughbreds were found to exhibit a statistically significant lower seroprevalence as a breed group when compared with other breeds sampled.Equine protozoal myeloencephalitis (EPM) is a disease that affects the central nervous system of the horse. The disease and its clinical presentation have been reviewed elsewhere. 4,7,8 In brief, the presenting clinical signs are variable but are related to central nervous system dysfunction that occurs in relation to infection and inflammation associated with a protozoal organism. Despite the fact that more than 1 type of protozoan has been associated with this disease, to date the most commonly associated protozoan is Sarcocystis neurona.Seroprevalence studies have been conducted in northern Colorado, Michigan, Ohio, Oregon, and southeastern Pennsylvania to estimate the rate of exposure of horses to S. neurona, relative to the geography of the USA. These reports have estimated overall statewide seroprevalences of 33.6%, 60%, 53.6%, 45%, and 45.3%, respectively. 2,3,10,12,13 To date, there are no published data pertaining to the seroprevalence of antibodies against S. neurona in the south-central USA.The purpose of this investigation was to estimate the seroprevalence of this antibody in the state of Oklahoma and to evaluate for relationships between seroprevalence and geographic location within the state and signalment of the test animals.Immunoblot analysis for anti-S. neurona antibody was performed as previously described 6 analysis by a commercial laboratory that offers immunoblot analysis for both serum and cerebrospinal fluid. a Data were recorded for each sample regarding county from which the sample was submitted, age of the horse, breed of the horse, and sex of the horse. Reactivity for proteins with relative mobilities of 14.5, 13, and 7 kD were considered positive indicators of exposure to S. neurona. Weak positive and low positive reactivities were regarded as positive because any reactivity indicates exposure to the organism. The overall seroprevalence was estimated from the results of the immunoblot analysis of 798 samples submitted. Statistical analysis was performed to determine the relationship of seroprevalence to age, breed, sex, and geographic quadrant within the state of Oklahoma. A computerized statistics program was used for all statistical calculations. b On the basis of 798 samples of equids and a 95% confidence interval, the overall statewide seroprevalence is estimated to be 89.2% Ϯ 2.1% (712/798). There were 792 samples for which geographic data regarding the origin of the sample were avai...

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Section: Seroprevalence Of Antibodies To Sarcocystis Neurona In Equidmentioning

confidence: 99%

Seroprevalence of Antibodies toSarcocystis Neuronain Equids Residing in Oklahoma

et al. 2003

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“…10,22 In brief, blots of the mWB were treated and incubated with fractionated bovine anti-S. cruzi IgG antibody diluted at 1:65 in blocking buffer from highly S. cruzi-positive cattle (Ͼl:200 in the IFAT) for 1.5 h before the addition of horse serum. 22 Merozoites of the SN-3 and MIH1 S. neurona isolates were used as antigens by the private and state laboratory, respectively.…”

Section: Western Blotsmentioning

confidence: 99%

“…A modified Western blot test (mWB) for detection of equine serum antibodies to S. neurona was recently developed. 22 The test was reported to have an improved specificity because blots were treated with bovine anti-Sarcocyctis cruzi antibodies before the addition of equine serum. On the basis of 6 S. neurona culture-positive horses (positive gold-standard) and 45 horses living in the eastern hemisphere and assumed never exposed to the parasite (negative gold-standard), the sensitivity and specificity of the mWB were 100% and 98%, respectively.…”

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confidence: 99%

“…On the basis of 6 S. neurona culture-positive horses (positive gold-standard) and 45 horses living in the eastern hemisphere and assumed never exposed to the parasite (negative gold-standard), the sensitivity and specificity of the mWB were 100% and 98%, respectively. 22 The indirect fluorescent antibody test (IFAT) has been evaluated and used for the diagnosis of protozoal diseases in many species, including horses, where the IFAT has been used to diagnose infections with Neospora, Babesia, and Trypanosoma species. 3,4,12,15,20,[27][28][29] However, cross-reactivity between S. neurona and other species of Sarcocystis infecting horses was thought to preclude the use of the IFAT for the diagnosis of EPM.…”

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confidence: 99%

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Comparison of a Serum Indirect Fluorescent Antibody Test with Two Western Blot Tests for the Diagnosis of Equine Protozoal Myeloencephalitis

et al. 2003

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Abstract.A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiveroperating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the area under the curves of the WB and the mWB (P ϭ 0.025 and P ϭ 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P Ͼ 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpreting weak positive WB results as positive test results, the sensitivities and 95% confidence intervals (CI) of all 3 tests were identical and equal to 88.9% (51.8-99.7%). The specificities and 95% CIs of the IFAT, WB, and mWB test were 100% (91-100%), 87.2% (72.6-95.7%), and 69.2% (52.4-83%), respectively. The overall accuracy of the IFAT was shown to be better than that of the WBs and, therefore, the test has potential for use in the diagnosis of EPM caused by Sarcocystis neurona.Equine protozoal myeloencephalitis (EPM) is a common neurological disease of horses in North America. 13,19 Sarcocystis neurona and Neospora hughesi are the protozoal parasites associated with EPM; 7,17 however, serologic surveys have indicated that exposure to S. neurona is more frequent. [1][2][3]21,26,27 The seroprevalence of S. neurona in US horses has been estimated to be between 34% and 60% in different parts of the country. 1,2,21,23,26,27 The Western blot (WB) test has been the standard diagnostic test used for serologic surveys and for the diagnosis of clinically suspected EPM, caused by S. neurona. The WB detects specific IgG antibodies in serum and spinal fluid (CSF) of horses infected or exposed to the parasite. 10 The overall sensitivity and specificity of the WB in CSF were estimated to be 87% and 56%, respectively. 5 In serum, the WB sensitivity and specificity were estimated as 80% and 38%, respectively, for horses with neurologic signs, and 88% and 56%, respectively, for horses without neurologic signs. 5 The low specificity of the test makes interpretation of a serologic positive test result difficult. To date, a negative WB test result on serum is

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“…Descriptions of testing options and reported test performance are shown in Table 1 71, 72, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93. All tests can be performed on serum or CSF, and none is considered a gold standard.…”

Section: Immunodiagnostic Testingmentioning

confidence: 99%

Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention

Reed

Furr

Howe

et al. 2016

Veterinary Internal Medicne

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Equine protozoal myeloencephalitis (EPM) remains an important neurologic disease of horses. There are no pathognomonic clinical signs for the disease. Affected horses can have focal or multifocal central nervous system (CNS) disease. EPM can be difficult to diagnose antemortem. It is caused by either of 2 parasites, Sarcocystis neurona and Neospora hughesi, with much less known about N. hughesi. Although risk factors such as transport stress and breed and age correlations have been identified, biologic factors such as genetic predispositions of individual animals, and parasite‐specific factors such as strain differences in virulence, remain largely undetermined. This consensus statement update presents current published knowledge of the parasite biology, host immune response, disease pathogenesis, epidemiology, and risk factors. Importantly, the statement provides recommendations for EPM diagnosis, treatment, and prevention.

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Improvement of Western Blot Test Specificity for Detecting Equine Serum Antibodies toSarcocystis Neurona

Cited by 48 publications

References 18 publications

Seroprevalence of Antibodies toSarcocystis Neuronain Equids Residing in Oklahoma

Seroprevalence of Antibodies toSarcocystis Neuronain Equids Residing in Oklahoma

Comparison of a Serum Indirect Fluorescent Antibody Test with Two Western Blot Tests for the Diagnosis of Equine Protozoal Myeloencephalitis

Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention

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