Journals / Türkiye Parazitoloji Dergisi / 2003 / Cilt: 27 - Sayı: 4

The diagnosis of Sarcocystis spp. in sheep using random amplified polymorphic DNA-PCR (RAPD-PCR)

Koyunlardaki Sarcocystis türlerinin random amplified polymorphic DNA-polimeraz zincir reaksiyonu (RAPD-PCR) ile teşhisi

Pages
255–259
Publication Date
2022-11-05
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

It was found that sarcocystosis is very common with the species S. tenella being dominant in sheep in the Konya Province.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:In this study, the RAPD-PCR (Random Amplified Polymorphic DNA-PCR) and the trypsin technique were used to investigate the presence and prevalence of the ovine Sarcocystis spp.as well as their molecular biology.
📊
Key Findings & Data:After optimization of the PCR conditions, three primers (OSA-Ol, OSA-02 and OSA-03) were selected to amplify DNA of the three parasite species.
💡
Conclusion & Impact:It was found that sarcocystosis is very common with the species S. tenella being dominant in sheep in the Konya Province.
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

In this study, the RAPD-PCR (Random Amplified Polymorphic DNA-PCR) and the trypsin technique were used to investigate the presence and prevalence of the ovine Sarcocystis spp.as well as their molecular biology. After optimization of the PCR conditions, three primers (OSA-Ol, OSA-02 and OSA-03) were selected to amplify DNA of the three parasite species. By generating band patterns with RAPD-PCR, it was possible to estimate the genetic relationship among the species in question. Among the pathogenic Sarcocystis spp., S. tenella and S. arieticanis that are grouped together are clearly distinct from S. gigantean, which forms a separate cluster. It was found that sarcocystosis is very common with the species S. tenella being dominant in sheep in the Konya Province. It was also found that the RAPD-PCR fingerprinting technique was more specific and sensitive than the trypsin technique.

Özet

Bu çalışmada, koyun Sarcocystis türlerinin yaygınlıklarının belirlenmesi ve moleküler biyolojilerinin araştırılması amacıyla Random Amplified Polymorphic DNA Polimeraz Zincir Reaksiyonu ve tripsin tekniği kullanılmıştır. Tür identifıkasyonu amacı ile S. gigantae için makroskobik ve mikroskobik kistlerden, S. arieticanis ve S. tenella için mikroskobik kistlerden yararlanılmıştır. DNA ekstraksiyonu için Fenol ekstraksiyon etanol prespitasyon metodu uygulanmıştır. Ayrıca, çalışmada Tripsin tekniği de uygulanarak, iki metodun sonuçları karşılaştırılmiştır. Koyunlardaki Sarcocystis türlerinin DNA'sını amplifiye etmek için farklı sekansa sahip üç primer (OSA-Ol, OSA-02 ve OSA-03) seçilmiştir. Random Amplified Polymorphic DNA Polimeraz Zincir Reaksiyonu (RAPD-PCR) ile amplifiye edilen band sayıları ve katsayılar sayesinde genetik ilişkinin değerlendirilmesi ile patojen türlerin (S. tenella, ve S. arieticanis), ayrı bir küme oluşturan S. gigantea'dan açık bir şekilde ayrıldığı tesbit edilmiştir. Sonuç olarak; Sarcocystis türlerinin Konya yöresi koyunlarında çok yaygın ve S. tenella'nm dominant tür olduğu belirlenmiş,RAPD-PCR fingerprinting tekniğinin teşhis açısından tripsin tekniğinden daha duyarlı ve özgül olduğu saptanmıştır.

🔖

Cite This Article

ALDEMİR, O. S., DİK, B. (2022). The diagnosis of Sarcocystis spp. in sheep using random amplified polymorphic DNA-PCR (RAPD-PCR). Türkiye Parazitoloji Dergisi, Cilt: 27 - Sayı: 4, 255–259. https://ajindex.com/articles/630938

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (17)

  1. Andrews CD. Dubey JP, Tenter AM, Webert DW, 1997. Toxoplasma gondii recombinant antigens H4, H11: use in ELISA for detection of toxoplasmosis in swine. Vet Parasitol, 70(1-3):1-11
  2. Blin N, Stafford DW, 1989. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acid Res, 3: 2303-2308
  3. Cerva L, Gut, J, 1983. Indirect Haemagglutination Reaction with antigen of Sarcocystis gigantea (Railliet, 1886) Ashford.Folia Parasitol (Praha), 30: 223-228
  4. Coote UG, 1990. Amplification of Nucleic Acids by the Polymerase Chain Reaction. Article, 4: 57-59
  5. Dubey JP, Desmonts G, 1987. Serological responses of equids fed Toxoplasma gondii oocysts. Equine Vet J, 19,(4): 337-339
  6. Dubey JP, Leek RG, Fayer R, 1986. Prevalence, transmission, pathogenicity of Sarcocystis gigantea of sheep. JAVMA, 188: 151-154
  7. Dubey JP, Specr CA, Fayer R, 1989 "Sarcocystosis of Animals, Man". CRC Press, Florida
  8. Ellis TJ, Luton K, Baverstock PR, Whitworth G, Tenter AM, Johnson AM, 1995. Phylogenetic relationships between Toxoplasma, Sarcocystis deduced from a comparison of 18S rDNA sequences. Parasitology, 110 (5): 521-528
  9. Göksu K, 1975. Koyunlarda sarcosporidiosisin yayılışı üzerine araştırmalar. İ.Ü.Vet Fak. Derg 1(1): 110-127
  10. Heckeroth AR, Tenter AM, 1999a. Development, validation of species- specific nested PCR for diagnosis of acute Sarcocystosis in sheep. Int J Parasitol, 29: 1331-1349
  11. Heckeroth AR, Tenter AM, 1999b. Comparison of immunological, molecular methods for the diagnosis of infections with pathogenic Sarcocystis species in sheep. Tokai J Ex Clin Med, 23: 293-302
  12. Holmdahl OJ, Mathson JG, Uggla A, Johansson KE. 1993. Oligonucleotide probes complementary to variable regions of 18S rRNA from Sarcocystis species. Mol Cell Probes, 7(6): 481-486
  13. Joachim AB, 1994. "Differentiation of ovine Sarcocystis species by RAPD-PCR fingerprinting". Institut fiir parasitologic der Tierarztlichen Hochschule, Hannover
  14. Joachim A, Tenter AM, Jeffries AC, Johnson AM, 1996. A RAPD-PCR derived marker can differentiate between pathogenic, non-pathogenic Sarcocystis species of sheep. Mol Cell Probes, 10(3): 165-172
  15. Kudi AC, Aganga AO, Ogbogu VC, Umoh JU, 1991 Prevalence of Sarcocystis species in sheep, goats. Vet Pays Trop, 44: 59-60
  16. Latif BMA, Al-Delemi JK, Mohammed BS, Al-Bayat SM, Al-Amiry AM, 1999. Prevalence of Sarcocystis spp. in meat-producing animals in Iraq. Vet Parasitol, 84: 85-90
  17. Leek RG, Fayer R, John