Journal of Parasitic Diseases

, Volume 33, Issue 1–2, pp 70–76 | Cite as

Standardization and application of indirect plate ELISA for immunodiagnosis of paramphistomosis in ruminants

  • Shivjot Kaur
  • L. D. Singla
  • S. S. Hassan
  • P. D. Juyal
Original Article


Indirect plate enzyme-linked immunosorbent assay was standardized and evaluated for its effectiveness in immunodiagnosis of paramphistomosis in experimental and clinical cases in sheep, goat, cattle and buffaloes by using somatic whole adult antigen of Paramphistomum epiclitum and Gastrothylax crumenifer. Plate enzymelinked immunosorbent assay (ELISA) was standardized using 2 μg/ml of antigen concentration with 1:200 and 1:1,000 of sera and conjugate dilution. Indirect Plate ELISA was able to demonstrate the antibody titre at different weeks postinfection in experimental sheep. Immune response at weekly interval varied in all the four experimental sheep. A paired t-test between two types of somatic antigens (P. epiclitum and G. crumenifer) showed that experimental sheep sera showed more affinity for homologous antigen as compared to heterologous antigen. A comparison of plate ELISA on suspected field sera and fecal samples examination by sedimentation method revealed that 77 samples were found to be positive by ELISA but only seven by fecal examination. Sensitivity of plate ELISA was found to be 85.71%, whereas specificity was 23.65% indicating that this test is quite sensitive for clinical cases; an early diagnosis, however, lacks specificity. In comparison to ELISA test the sensitivity and specificity of fecal examination were 7.79 and 88%, respectively.


Indirect plate ELISA Immunodiagnosis Paramphistomum epiclitum Somatic antigen 


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  1. Alabay M (1981) Investigation on the diagnosis of Paramphistomum cervi infections in immunoperoxidase and IFAT technique in sheep. Ankara Universities Veterier Facultes: Dergisi 21: 72–88Google Scholar
  2. Anuracpreeda P, Wanichanon C, Sobhon P (2008) Paramphistomum cervi: antigenic profile of adults as recognized by infected cattle sera. Exptl Parasitol 118:203–207CrossRefGoogle Scholar
  3. Baldrey FSH (1906) Some problems in sheep diseases. J Trop Vet Sci 1:387–409Google Scholar
  4. Bawa HS (1939) Intestinal paramphistomiasis of sheep in Sind (a preliminary report). Indian J Vet Sci Anim Husb 9:425–429Google Scholar
  5. Boch J, Schmid K, Ruckrich HU, Erich E, Keller B, Weiland G, Gobel E (1983) Stomach fluke infection (Paramphistomum cervi) infection of ruminants. Berliner-und-Munchener-Tierarztliche-Wochenschrift 96:338–346Google Scholar
  6. Chhabra RC, Gill BS, Dutt SC (1978) Paramphistomiasis in sheep and goats in Punjab state and its treatment. Indian J Parasitol 2:43–45Google Scholar
  7. Dutt SC (1980) Paramphistomes and paramphistomiasis of domestic ruminants in India. Singh A (Ed.), Punjab Agric University, LudhianaGoogle Scholar
  8. Hafeez MD (2003) Helminth parasites of public health importance-Trematodes. J Parasit Dis 27:69–75Google Scholar
  9. Hafeez MD, Avastthi BL (1987) Incidence of ruminal amphistomes of certain domestic ruminants in and around Gujrat. Gujvet 15:18–20Google Scholar
  10. Hassan SS, Kaur K, Joshi K, Juyal PD (2005) Epidemiology of paramphistomosis in domestic ruminants in different districts of Punjab and other adjoining areas. J Vet Parasitol 19:43–46Google Scholar
  11. Horak IG (1971) Paramphistomiasis of domestic ruminants. Adv Parasitol 9:33–72CrossRefPubMedGoogle Scholar
  12. Hudson L, Hay FC (1989) Practical Immunology. 3rd edition, Blackwell Scientific Publications, Oxford, London, pp 507Google Scholar
  13. Katiyar RD, Varshney TR (1963) Amphistomiasis in sheep and goats in Uttar Pradesh. Indian J Vet Sci Anim Husb 33:94–98Google Scholar
  14. Kaur K, Hassan SS, Kaur P, Juyal PD (2004) Indirect Dot-ELISA for anti-Paramphistomum epiclitum antibodies detection. J Vet Parasitol 18(1):73–74Google Scholar
  15. Keller B (1983) Application of the serological techniques ELISA and IFAT to Paramphistomum cervi infection in sheep. Inaugural Dissertation, Tierarztliche Fakultate, Ludwing-Maximilians-Universitat Munclen, pp 56Google Scholar
  16. Lowry OH, Rosenbrough NJ, Far AL, Randael RJ (1951) Protein measurement with Folin phenol reagents. J Biol Chem 193: 265–275PubMedGoogle Scholar
  17. Malviya HC, Prasad A, Varma TK, Dwivedi P (1989) On the collection of Paramphistomum cervi metacercariae and their infectivity to lambs. J Vet Parasitol 3:53–55Google Scholar
  18. Matto NR, Bali HS (1991) Incidence of helminths in Indian buffaloes. Indian J Parasitol 15:11–12Google Scholar
  19. Pande BK, Misra SC (1980) Observations on the etiology, clinical pathology and chemotherapy of immature amphistomiasis in buffalo calves. Indian J Anim Hlth 19:131–135Google Scholar
  20. Pande PG (1935) Acute amphistomiasis of cattle in Assam (A preliminary report). Indian J Vet Sci Anim Husb 5:364–375Google Scholar
  21. Phiri AM, Chota A, Phiri IK (2007) Seasonal pattern of bovine amphistomosis in traditionally reared cattle in the Kafue and Zambezi catchment areas of Zambia. Trop Anim Health Prod 37:97–102CrossRefGoogle Scholar
  22. Prasad A, Varma TK (1999) On the prevalence and community dominance among paramphistomes infecting domestic ruminants. J Vet Parasitol 13:129–133Google Scholar
  23. Soulsby EJL (1982) Helminths, Arthropods and Protozoa of Domesticated Animals. 7th edition, Bailliere-Tindall, LondonGoogle Scholar
  24. Tariq KA, Chishti MZ, Ahmad F, Shawl AS (2008) Epidemiological study of Paramphistomum infection in goats-Kashmir Valley. World J Agric Sci 4:61–64Google Scholar
  25. Wang CR, Qiu JH, Zhu XQ, Han XH, Ni HB, Zhao JP, Zhou QM, Zhang HW, Lun ZR (2006) Survey of helminthes in adult sheep in Heilongjiang Province, People’s Republic of China. Vet Parasitol 140:378–382CrossRefPubMedGoogle Scholar

Copyright information

© Indian Society for Parasitology 2009

Authors and Affiliations

  • Shivjot Kaur
    • 1
  • L. D. Singla
    • 1
  • S. S. Hassan
    • 1
  • P. D. Juyal
    • 1
  1. 1.Department of Veterinary Parasitology, College of Veterinary ScienceGuru Angad Dev Veterinary and Animal Sciences UniversityLudhianaIndia

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