1. bookVolume 64 (2014): Issue 1 (March 2014)
Journal Details
License
Format
Journal
eISSN
1820-7448
First Published
25 Mar 2014
Publication timeframe
4 times per year
Languages
English
Open Access

Characteristic band pattern in western blots for specific detection of anti-Trichinella spiralis antibodies in different host species

Published Online: 25 Mar 2014
Volume & Issue: Volume 64 (2014) - Issue 1 (March 2014)
Page range: 33 - 43
Journal Details
License
Format
Journal
eISSN
1820-7448
First Published
25 Mar 2014
Publication timeframe
4 times per year
Languages
English
Abstract

Western blot (Wb) is considered to be the gold standard test for Trichinella infection serology, since this method allows specific Trichinella antigens to be distinguished from cross-reactive antigens. This is not the case with widely used antibody assay techniques - indirect immunofluorescence and ELISA - which are sensitive, but subject to crossreactions that make the interpretation of weakly positive results difficult. Application of Trichinella spiralis muscle larvae excretory-secretory (ES) antigens for the specific antibody detection in ELISA resulted in improved specificity compared to that of crude worm extract that was previously in use, but since production of ES has not yet been standardized, differences among laboratories occur. For this reason, the Wb profile of serum samples from different T. spiralis infected host species: human, horse, swine and dog, was investigated in the Serbian National Reference Laboratory for Trichinellosis (NRLT). The common feature of the obtained Wb profiles was the appearance of a triad of bands with molecular masses (Mw) of 45, 49, and 53 kDa. The very same triad was recognized by a monoclonal antibody (mAb) 7C2C5 specific for an immunodominant epitope unique to the muscle larvae stage of all species in the genus Trichinella. Inhibition studies confirmed that mAb and anti-Trichinella antibodies from sera competed for the same parasite epitope. Based on the obtained results, the NRLT introduced the recognition of the above mention triad as the basis for specific anti-Trichinella antibodies detection in the sera of infected hosts.

Keywords

1. Murell KD, Pozio E: Worldwide occurrence and impact of human trichinellosis, 1986-2009. Emerg Infect Disease 2011, 17:2194-2202.10.3201/eid1712.110896Search in Google Scholar

2. Cuperlovic K, Sofronic-Milosavljevic Lj, Hristovic V: Trichinellosis - immunodiganosis and disease. In: Trichinellosis (G.Ortega-Pierres, H.R.Gamble, F. van Knapen, D. Wakelin, eds.) Centro de Investigacion y Estudios Avanzados IPN, D.F. Mexico, Mexico; 1996, 513-518.Search in Google Scholar

3. Dell A, Haslam SM, Morris HR, Khoo KH: Immunogenic glycoconjugates implicated in parasitic nematode diseases. Biochem Biophys Acta 1999, 1445:353-362.10.1016/S0925-4439(99)00064-2Search in Google Scholar

4. Bruschi F, Chiumiento L: Immunomodulation in Trichinellosis: Does Trichinella Really Escape the Host Immune System? Endocr Metab Immune Disord Drug Targets 2012, 12:4-15.10.2174/18715301279927908122214331Search in Google Scholar

5. Sofronic-Milosavljevic Lj, Djordjevic M, Plavsic B, Grgic B: Trichinella infection in Serbia in the fi rst decade of the twenty-fi rst century. Vet Parasitol 2013, 194:136-138.10.1016/j.vetpar.2013.01.03923453823Search in Google Scholar

6. Cuperlovic K, Djordjevic M: Trichinella et Trichinellosis. (Turbatovic L, ed.), Institut za higijenu i tehnologiju mesa, Beograd; 2003.Search in Google Scholar

7. Cuperlovic K, Djordjevic M, Pavlovic S: Reemergence of trichinellosis in southeast Europe due to political and economic changes. Vet Parasitol 2005, 132:159-166.10.1016/j.vetpar.2005.05.04715979803Search in Google Scholar

8. Pozio E, Rinaldi L, Marruci G, Musella V, Galati F, Cringoli G, Boireau P, La Rosa G: Host and habitats of Trichinella spiralis and Trichinella britovi in Europe. Int J Parasitol 2009, 39:71-79.10.1016/j.ijpara.2008.06.00618708065Search in Google Scholar

9. Cvetkovic J, Teodorovic V, Marucci G, Vasilev D, Vasilev S, Cirovic D, Sofronic-Milosavljevic Lj: First report on Trichinella britovi in Serbia. Acta Parasitol 2011, 56:232-235.10.2478/s11686-011-0022-1Search in Google Scholar

10. Zivojinovic M, Sofronic-Milosavljevic L, Cvetkovic J, Pozio E, Interisano M, Plavsic B, Radojicic S, Kulisic Z: Trichinella infections in different host species of an endemic district of Serbia. Vet Parasitol 2013, 194:136- 138.10.1016/j.vetpar.2013.01.039Search in Google Scholar

11. Cuperlovic K, Djordjevic M, Pavlovic S, Sofronic Milosavljevic L: Present status of trichinellosis in Yugoslavia: Serbia. Parasite 2001, 8:95-97.10.1051/parasite/200108s209511484397Search in Google Scholar

12. Murrell KD, Djordjevic M, Cuperlovic K, Sofronic Lj, Savic M, Djordjevic M, Damjanovic S: Epidemiology of Trichinella infection in the horse: the risk from animal product feeding practices. Vet Parasitol 2004, 123:223-233.10.1016/j.vetpar.2004.06.00815325048Search in Google Scholar

13. Gamble HR, Pozio E, Bruschi F, Nockler K, Kapel CMO, Gajadhar AA: International Commission on Trichinellosis: recommendations on the use of serological tests for the detection of Trichinella infection in animals and man. Parasite 2004, 11:3-1310.1051/parasite/20041113Search in Google Scholar

14. Gamble HR: Detection of trichinellosis in pigs by artifi cial digestion and enzyme immunoassay. J Food Protect 1996, 59:295-298.10.4315/0362-028X-59.3.295Search in Google Scholar

15. Noeckler K, Pozio E, Voigt WP, Heidrich J: Detection of Trichinella infection in food animals. Vet Parasitol 2000, 93:335-350.10.1016/S0304-4017(00)00350-2Search in Google Scholar

16. Gottstein B, Pozio E, Nockler K: Epidemiology, Diagnosis, Treatment, and Control of Trichinellosis. Clin Microbiol Rev 2009, 22:127-145.10.1128/CMR.00026-08Search in Google Scholar

17. Gamble HR, Bessonov AS, Cuperlovic K, Gajadhar AA, Van Knapen F, Noeckler K, Schenone H, Zhu X: International commission on trichinellosis: recommendations on methods for control of Trichinella in domestic and wild animals intended for human consumption. Vet Parasitol 2000, 93:393-408.10.1016/S0304-4017(00)00354-XSearch in Google Scholar

18. Gamble HR, Graham CE: Monoclonal antibody-purifi ed antigen for the immunodiagnosis of trichinosis. Am Vet Med Res 1984, 45:67-74.Search in Google Scholar

19. Sofronic-Milosavljevic Lj, Cuperlovic K, Cokic Z: Tumor originated antibodies from tumor patients recognize Trichinella spiralis antigens. In: Trichinellosis (G.Ortega-Pierres, H.R.Gamble, F. van Knapen, D. Wakelin, eds.) Centro de Investigacion y Estudios Avanzados IPN, D.F. Mexico, Mexico; 1996, 233-238.Search in Google Scholar

20. Robert F, Weil B, Kassis N, Dupouy-Camet J: Investigation of immunofl uorescence crossreactions against Trichinella spi ralis by Western blot (immunoblot) analysis. Clin Diagn Lab Immunol 1996, 5:575-577.10.1128/cdli.3.5.575-577.19961704098877138Search in Google Scholar

21. Radovic I, Gruden-Movsesijan A, Ilic N, Mostarica-Stojkovic M, Sofronic-Milosavljevic Lj: Trichinella spiralis shares epitopes with human autoantigens. Mem Instit Oswaldo Cruz 2012, 107:503-509.10.1590/S0074-0276201200040001022666861Search in Google Scholar

22. Linder E, Thors C, Lundin L, Ljungström I, Farah S, Hagi H, Dias F: Schistosome antigen gp50 is responsible for serological cross-reactivity with Trichinella spiralis. J Parasitol 1992, 78:999-1005.10.2307/3283220Search in Google Scholar

23. De la Rosa JL, Alcantara P, Correa D: Investigation of cross-reactions against Trichinella spiralis antigens by enzyme-linked immunosorbent assay and enzyme-linked immunoelectrotransfer blot assay in patients with various diseases. Clin Diagn Lab Immunol 1995, 2:122-4.10.1128/cdli.2.1.122-124.19951701137719905Search in Google Scholar

24. Intapan PM, Maleewong W, Sukeepaisarnjaroen W, Morakote N: Potential use of Trichinella spiralis antigen for serodiagnosis of human capillariasis philippinensis by immunoblot analysis. Parasitol Res 2006, 98:227-231.10.1007/s00436-005-0070-216333664Search in Google Scholar

25. Robinson M, Gustrad TR, Erickson MR: Immunological interactions between Trichinella spiralis and Heligmosomoides polygyrus: cross reactivity between muscle larvae and antibodies raised to unrelated antigens. Int J Parasitol 1997, 27:865-872.10.1016/S0020-7519(97)00049-0Search in Google Scholar

26. Møller LN, Krause TG, Koch A, Melbye M, Kapel CM, Petersen E: Human antibody recognition of Anisakidae and Trichinella spp. in Greenland. Clin Microbiol Infec 2007, 13:702-708.10.1111/j.1469-0691.2007.01730.xSearch in Google Scholar

27. Ivanoska D, Cuperlovic K, Gamble HR, Murrell KD: Comparative effi cacy of antigen and antibody detection tests for human trichinellosis. J Parasitol 1989, 75:38-41.10.2307/3282932Search in Google Scholar

28. Constantino SN, Malmassari SL, Dalla Fontana ML, Diamante MA, Venturiello SM: Diagnosis of human trichinellosis: pitfalls in the use of a unique immunoserological technique. Parasite 2001, 8:144-146.10.1051/parasite/200108s2144Search in Google Scholar

29. Pozio E, Sofronic-Milosavljevic Lj, Gomez Morales MA, Boireau P, Nockler K: Evaluation of ELISA and western blot analysis using three antigens to detect anti-Trichinella IgG in horses. Vet Parasitol 2002, 108:163-178.10.1016/S0304-4017(02)00185-1Search in Google Scholar

30. Sofronic-Milosavljevic Lj, Ilic N, Djordjevic M, Savic M, Gruden-Movsesijan A, Cuperlovic K, Murell KD: Anti-Trichinella antibodies detected in chronically infected horses by IFA and Western blot, but not by ELISA. Vet Parasitol Supp 2005, 132:107-11.10.1016/j.vetpar.2005.05.037Search in Google Scholar

31. Soule C, Dupouy-Camet J, Georges P, Ancelle T, Gillet JP, Vaissaire J, Delvigne A, Plareau E: Experimental trichinellosis in horse: Biological and parasitological evaluation.Search in Google Scholar

Vet Parasitol 1989, 31:19-36.10.5840/gps19893614Search in Google Scholar

32. Yepez-Mulia L, Arriaga C, Viveros N, Adame A, Benitez E, Ortega-Pierres MG: Detection of Trichinella infection in slaughter horses by ELISA and western blot analysis. Vet Parasitol 1999, 81:57-68.10.1016/S0304-4017(98)00208-8Search in Google Scholar

33. Boireau P, Valle I, Roman T, Perret C, Mingyuan L, Gamble HR, Gajadhar A: Trichinella in horses: a low frequency infection with high human risk. Vet Parasitol 2000, 93:309-320.10.1016/S0304-4017(00)00348-4Search in Google Scholar

34. Hill DE, Forbes L, Kramer M, Gajadhar A, Gamble HR:. Larval viability and serological response in horses with long-term Trichinella spiralis infection. Vet Parasitol 2007, 146:107-116.10.1016/j.vetpar.2007.02.01117386976Search in Google Scholar

35. Yera H, Andiva S, Perret C, Limonne D, Boireau P, Dupouy-Camet J: Development and evaluation of a western blot kit for diagnosis of human trichinellosis. Clin Diagn Lab Immunol 2003, 10:793-796.10.1128/CDLI.10.5.793-796.2003Search in Google Scholar

36. Gomez-Morales MA, Ludovisi A, Amati M, Blaga R, Zivojinovic M, Ribicich M, Pozio E: A distinctive Western blot pattern to recognize Trichinella infections in humans and pigs. Int J Parasitol 2012, 42:1017-1023. 10.1016/j.ijpara.2012.08.00322995743Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo