FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Am J Vet Res / Sequence analysis and polymerase chain reaction amplificatio...
American journal of veterinary research1996; 57(7); 975-981;

Sequence analysis and polymerase chain reaction amplification of small subunit ribosomal DNA from Sarcocystis neurona.

Abstract: To identify Sarcocystis neurona-specific DNA sequences in the nuclear small subunit ribosomal RNA (nss-rRNA) gene that could be used to distinguish S neurona from other closely related protozoal parasites, and to evaluate a polymerase chain reaction (PCR) test, using broad based primers and a unique species-specific probe on CSF for detection of S neurona in equids. Methods: Sequencing of the nuclear small subunit ribosomal RNA gene from a new S neurona isolate (UCD 1) was performed. The sequence was compared with that of other closely related Sarcocystidae parasites. From this sequence, conserved DNA sequence primers were selected and an oligonucleotide probe was designed to hybridize with a unique region of the S neurona gene. For clinical evaluation, horses were considered test positive for S neurona infection on the basis of immunohistochemical detection of the parasites in the CNS. Results: Sensitivity of this PCR and probe-based detection system was approximately 1 to 5 merozoites. Cerebrospinal fluid from 2 of 5 horses with histologic lesions consistent with equine protozoal myeloencephalitis were PCR- and probe-positive in a blind test of this procedure, and all uninfected horses were test negative. Conclusions: This PCR-based system is a useful method of confirming S neurona in CSF and has the advantage of facilitating detection of other apicomplexan protozoans that may be infective for horses. The usefulness of this test is limited by the presence of parasites free in the CSF of clinically affected horses.
Get Full Text
Publication Date: 1996-07-01 PubMed ID: 8807006
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.
LinkedInCopy
  • Comparative Study
  • 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 study examines the process of identifying specific DNA sequences in Sarcocystis neurona and using them to differentiate it from similar parasites through polymerase chain reaction (PCR) test. The research utilizes the test on equines to assess its sensitivity and efficacy in a clinical setting.

Methodology

  • The researchers conducted a sequence analysis of a newly isolated Sarcocystis neurona (UCD 1) strain. This was done on the nuclear small subunit ribosomal RNA (nss-rRNA) gene.
  • This sequence was then compared with those of other closely related parasites from the Sarcocystidae family to identify unique sequences.
  • Using the unique sequence identified, conserved DNA sequence primers were selected and an oligonucleotide probe was designed. This probe would hybridize with the unique region of the S neurona gene.
  • For clinical testing, horses were treated as test positives for S neurona infection based on immunohistochemical detection of the parasites in their central nervous system (CNS).

Results

  • The sensitivity of the PCR and probe-based detection system ranged from approximately 1 to 5 merozoites.
  • Among five horses with histologic lesions consistent with equine protozoal myeloencephalitis, two tested positive when blind tested with this procedure. All uninfected horses tested negative.

Conclusion

  • The researchers concluded that the PCR-based system is a useful tool to confirm S neurona in cerebrospinal fluid (CSF).
  • Furthermore, this system can also facilitate the detection of other apicomplexan protozoans that may infect horses.
  • However, the test’s utility is limited by the presence of parasite-free CSF in clinically affected horses.

Cite This Article

APA
Marsh AE, Barr BC, Madigan J, Lakritz J, Conrad PA. (1996). Sequence analysis and polymerase chain reaction amplification of small subunit ribosomal DNA from Sarcocystis neurona. Am J Vet Res, 57(7), 975-981.

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 57
Issue: 7
Pages: 975-981

Researcher Affiliations

Marsh, A E
  • Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis 95616, USA.
Barr, B C
    Madigan, J
      Lakritz, J
        Conrad, P A

          MeSH Terms

          • Animals
          • Base Sequence
          • Cerebrospinal Fluid / parasitology
          • DNA, Protozoan / chemistry
          • DNA, Ribosomal / chemistry
          • Encephalomyelitis / diagnosis
          • Encephalomyelitis / parasitology
          • Encephalomyelitis / veterinary
          • Horse Diseases
          • Horses
          • Molecular Sequence Data
          • Polymerase Chain Reaction / methods
          • Reference Values
          • Reproducibility of Results
          • Sarcocystis / classification
          • Sarcocystis / isolation & purification
          • Sarcocystosis / diagnosis
          • Sarcocystosis / veterinary
          • Sensitivity and Specificity

          Citations

          This article has been cited 7 times.
          1. Enriquez CK, Morrow JK, Graves A, Johnson A. Evaluation of real-time polymerase chain reaction for the diagnosis of protozoal myeloencephalitis in horses using cerebrospinal fluid.. J Vet Intern Med 2023 Sep-Oct;37(5):1893-1898.
            doi: 10.1111/jvim.16826pubmed: 37549306google scholar: lookup
          2. Britton AP, Bidulka J, Scouras A, Schwantje H, Joseph T. Fatal hepatic sarcocystosis in a free-ranging grizzly bear cub associated with Sarcocystis canis-like infection.. J Vet Diagn Invest 2019 Mar;31(2):303-306.
            doi: 10.1177/1040638719826627pubmed: 30698508google scholar: lookup
          3. Kalantari N, Khaksar M, Ghaffari S, Hamidekish SM. Molecular Analysis of Sarcocystis Spp. Isolated from Sheep (Ovis aries) in Babol Area, Mazandaran Province, Northern Iran.. Iran J Parasitol 2016 Jan-Mar;11(1):73-80.
            pubmed: 27095971
          4. Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).. Vet Parasitol 2015 Apr 15;209(1-2):1-42.
            doi: 10.1016/j.vetpar.2015.01.026pubmed: 25737052google scholar: lookup
          5. Miller MA, Conrad PA, Harris M, Hatfield B, Langlois G, Jessup DA, Magargal SL, Packham AE, Toy-Choutka S, Melli AC, Murray MA, Gulland FM, Grigg ME. A protozoal-associated epizootic impacting marine wildlife: mass-mortality of southern sea otters (Enhydra lutris nereis) due to Sarcocystis neurona infection.. Vet Parasitol 2010 Sep 20;172(3-4):183-94.
            doi: 10.1016/j.vetpar.2010.05.019pubmed: 20615616google scholar: lookup
          6. Rejmanek D, Miller MA, Grigg ME, Crosbie PR, Conrad PA. Molecular characterization of Sarcocystis neurona strains from opossums (Didelphis virginiana) and intermediate hosts from Central California.. Vet Parasitol 2010 May 28;170(1-2):20-9.
            doi: 10.1016/j.vetpar.2009.12.045pubmed: 20226596google scholar: lookup
          7. Wendte JM, Miller MA, Nandra AK, Peat SM, Crosbie PR, Conrad PA, Grigg ME. Limited genetic diversity among Sarcocystis neurona strains infecting southern sea otters precludes distinction between marine and terrestrial isolates.. Vet Parasitol 2010 Apr 19;169(1-2):37-44.
            doi: 10.1016/j.vetpar.2009.12.020pubmed: 20071081google scholar: lookup
          Subscribe to our newsletter
          Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.