Effects of blood contamination of cerebrospinal fluid on western blot analysis for detection of antibodies against Sarcocystis neurona and on albumin quotient and immunoglobulin G index in horses.
Abstract: To determine effects of blood contamination on western blot (WB) analysis of CSF samples for detection of anti-Sarcocystis neurona antibodies, and on CSF albumin and IgG concentrations, albumin quotient (AQ), and IgG index in horses. Methods: Prospective in vitro study. Methods: Blood with various degrees of immunoreactivity against S neurona was collected from 12 healthy horses. Cerebrospinal fluid without immunoreactivity against S neurona was harvested from 4 recently euthanatized horses. Methods: Blood was serially diluted with pooled nonimmunoreactive CSF so that final dilutions corresponded to 10(-3) to 100 microliters of blood/ml CSF, and WB analysis was performed on contaminated CSF samples. Number of RBC, albumin and IgG concentrations, AQ, and IgG index were also determined. Results: Antibodies against S neurona were detected in CSF contaminated with 10(-3) microliters of strongly immunoreactive blood/ml. In CSF samples contaminated with 10 microliters of blood/ml, AQ remained within reference range. Volume of blood required to increase IgG index varied among blood samples and was primarily influenced by serum IgG concentrations. Number of RBC in contaminated samples was correlated with volume of blood added, but not with degree of immunoreactivity detected in contaminated CSF samples. Conclusions: During collection of CSF from horses, contamination with blood may introduce serum antibodies against S neurona at concentrations sufficient for detection by WB analysis, thus yielding false-positive results. When blood is moderately or strongly immunoreactive, the amount of contaminating albumin may be small enough as to not increase AQ above reference range. In these cases, AQ and IgG index should be interpreted with caution.
Publication Date: 1999-07-09 PubMed ID: 10397068
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research paper discusses a study conducted to determine the impact of blood contamination on the analysis of horse cerebrospinal fluid (CSF) samples for detecting antibodies against Sarcocystis neurona using western blot (WB) analysis. The study also evaluates how blood contamination affects CSF albumin and immunoglobulin G (IgG) levels, along with the albumin quotient (AQ) and the IgG index.
Study Methods
- The researchers used a prospective in vitro study design for this investigation.
- Blood was collected from healthy horses and was characterised by varying degrees of immunoreactivity against S. neurona.
- CSF without any immunoreactivity against S. neurona was taken from four horses that were recently euthanized.
- The blood sample was gradually diluted with pooled nonimmunoreactive CSF. The final dilutions translated to a range from 10(-3) to 100 microliters of blood per milliliter of CSF.
- WB analysis was performed on the contaminated CSF samples. Along with this, the number of red blood cells (RBC), albumin and IgG concentrations, AQ, and IgG index were also determined.
Results
- The results revealed that even CSF samples contaminated with 10(-3) microliters of highly immunoreactive blood per milliliter could still detect antibodies against S. neurona.
- CSF samples contaminated with 10 microliters of blood per milliliter maintained the AQ within the reference range.
- The volume of blood required to increase the IgG index varied among samples, meaning it was primarily influenced by the serum IgG concentrations rather than the volume of contaminated blood.
- The RBC count in contaminated samples was correlated with the volume of blood added, but not with the degree of immunoreactivity detected in the contaminated CSF samples.
Conclusions
- The research concludes that blood contamination during the collection of CSF from horses could introduce serum antibodies against S. neurona at concentrations high enough for detection by WB analysis, potentially leading to false-positive results.
- When the blood is moderately or highly immunoreactive, the amount of contaminating albumin might be minimal enough to not elevate the AQ above the reference range. Therefore, in such cases, the AQ and IgG index should be interpreted cautiously.
Cite This Article
APA
Miller MM, Sweeney CR, Russell GE, Sheetz RM, Morrow JK.
(1999).
Effects of blood contamination of cerebrospinal fluid on western blot analysis for detection of antibodies against Sarcocystis neurona and on albumin quotient and immunoglobulin G index in horses.
J Am Vet Med Assoc, 215(1), 67-71.
Publication
Researcher Affiliations
- Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square 19348-1692, USA.
MeSH Terms
- Albumins / cerebrospinal fluid
- Animals
- Antibodies, Protozoan / blood
- Antibodies, Protozoan / cerebrospinal fluid
- Blotting, Western / veterinary
- Encephalomyelitis / cerebrospinal fluid
- Encephalomyelitis / diagnosis
- Encephalomyelitis / veterinary
- Erythrocyte Count / veterinary
- False Positive Reactions
- Horse Diseases / cerebrospinal fluid
- Horse Diseases / diagnosis
- Horses
- Immunoglobulin G / blood
- Immunoglobulin G / cerebrospinal fluid
- Prospective Studies
- Sarcocystis / immunology
- Sarcocystosis / cerebrospinal fluid
- Sarcocystosis / diagnosis
- Sarcocystosis / veterinary
- Specimen Handling / standards
- Specimen Handling / veterinary
Citations
This article has been cited 5 times.- Puerto-Parada M, Arango-Sabogal JC, Bilodeau MÈ, Bédard C, Francoz D, Desrochers A, Nichols S, Fecteau G. Interpretation of cerebrospinal fluid analysis from recumbent cows using different thresholds of red blood cell count. J Vet Intern Med 2022 Sep;36(5):1837-1842.
- Chidlow H, Giguère S, Camus M, Wells B, Howerth E, Berghaus R, McConachie Beasley E. Comparison of 2 collection methods for cerebrospinal fluid analysis from standing, sedate adult horses. J Vet Intern Med 2020 Mar;34(2):972-978.
- Reed SM, Furr M, Howe DK, Johnson AL, MacKay RJ, Morrow JK, Pusterla N, Witonsky S. Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention. J Vet Intern Med 2016 Mar-Apr;30(2):491-502.
- Braun U, Attiger J, Brammertz C. Ultrasonographic examination of the spinal cord and collection of cerebrospinal fluid from the atlanto-occipital space in cattle. BMC Vet Res 2015 Sep 2;11:227.
- Steinberg TA, Boettcher IC, Matiasek K, Hirschvogel K, Hartmann K, Kunz A, Fischer A. Use of albumin quotient and IgG index to differentiate blood- vs brain-derived proteins in the cerebrospinal fluid of cats with feline infectious peritonitis. Vet Clin Pathol 2008 Jun;37(2):207-16.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
