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The potential use of 29 kDa protein as a marker of pathogenicity and diagnosis of symptomatic infections with Blastocystis hominis

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Abstract

The present study was performed to characterize the protein profiles of Blastocystis hominis isolates from symptomatic and asymptomatic individuals by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from symptomatic and asymptomatic patients. The presence of immunogenic bands associated with pathogenicity or of diagnostic potentials was also evaluated. The study comprised 80 individuals classified into four groups, 20 each: symptomatic blastocystosis (G1), asymptomatic blastocystosis (G2), other parasitic infections (G3), and healthy control subjects (G4). SDS-PAGE analysis of individual antigens form symptomatic and asymptomatic B. hominis isolates revealed similar and distinctive antigenic bands with significant differences in two high (123.5 and 112.3 kDa) and few low molecular weight bands (48.5, 38, 42.3, and 35.5 kDa). Immunoblotting was performed using symptomatic and asymptomatic antigen pools with sera of the four studied groups. It was found that anti-B. hominis IgG reacted with nine protein bands ranging from 100 to 18 kDa of the symptomatic antigen pool. There was a significant difference between G1 and G2 in the recognition of 64, 56, 38, and 29 kDa antigen bands. Also, anti-B. hominis IgG reacted with five protein bands ranging from 56 to 12 kDa of asymptomatic antigen pool. There was a significant difference between G1 and G2 in the recognition of 29 kDa antigen band. These findings suggest the potential use of the 29-kDa antigen as marker of pathogenicity and implicate its use in the diagnosis and differentiation between symptomatic and asymptomatic blastocystosis.

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References

  • Boreham PFL, Stenzel DJ (1993) Blastocystis in humans and animals: morphology, biology, and epizootiology. Adv Parasitol 32:1–70

    Article  PubMed  CAS  Google Scholar 

  • Boreham PFL, Upcroft JA, Dunn LA (1992) Protein and DNA evidence for two demes of Blastocystis hominis from humans. Int J Parasitol 22:49–53

    Article  PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Chen J, Vaudry WL, Kowalewska K, Wenman W (1987) Lack of serum immune response to Blastocystis hominis. Lancet 2:1021, Letter

    Article  PubMed  CAS  Google Scholar 

  • Chen XQ, Singh M, Ho LC, Tan SW, Yap EH (1999) Characterization of protein profiles and cross-reactivity of Blastocystis antigens by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis. Parasitol Res 85:343–346

    Article  PubMed  CAS  Google Scholar 

  • Diamond LS (1983) Lumen dwelling protozoa: Entamoeba, trichomonads and Giardia. In: Jensen JB (ed) In vitro cultivation of protozoan parasites. CRC, Boca Raton, Florida, pp 65–109

    Google Scholar 

  • Elwakil HS, Talaat RM (2009) Genetic analysis of Blastocystis hominis isolated from symptomatic and asymptomatic human hosts in Egypt. J Egypt Soc Parasitol 39(1):99–109

    PubMed  Google Scholar 

  • Elwakil HS, Hewedi IH (2010) Pathogenic potential of Blastocystis hominis in laboratory mice. Parasitol Res 107:685–689

    Article  PubMed  Google Scholar 

  • Garcia LS, Bruchner DA (1993) Macroscopic and microscopic examination of fecal specimens, in “Diagnostic Medical Parasitology” 2nd edition. Pub. The American Society of Microbiology, Washington, D.C. Chapter 26, pp 501–540

  • Hegazy MM, Maklouf LM, El Hamshary EM, Dawoud HA, Eida AM (2008) Protein profile and morphometry of cultured human Blastocystis hominis from children with gastroenteritis and healthy ones. JEgypt Soc Parasitol 38(2):453–464

    CAS  Google Scholar 

  • Hussain R, Jaferi W, Zuberi S, Baqai R, Abrar W, Ahmed A, Zaman V (1997) Significantly increased IgG2 subclass antibody levels to Blastocystis hominis in patients with irritable bowel syndrome. Am JTrop Med Hyg 56:301–305

    CAS  Google Scholar 

  • Hussein EM, Hussein AM, Eida MM, Atwa MM (2008) Pathophysiological variability of different genotypes of human Blastocystis hominis Egyptian isolates in experimentally infected rats. Parasitol Res 102:853–860

    Article  PubMed  Google Scholar 

  • Init I, Mak JW, Top S, Zulhainan Z, Prummongkol S, Nissapatorn V, Wan-Yussof WS, Anuar AK (2003) Polypeptides associated with in vitro cyst formation of Blastocystis hominis. Southeast Asian J Trop Med Public Health 34:727–732

    PubMed  CAS  Google Scholar 

  • Kaneda Y, Horiki N, Cheng XJ, Tachibana H, Tsutsumi Y (2000) Serologic response to Blastocystis hominis infection in asymptomatic individuals. Tokai J Exp Clin Med 25:51–56

    PubMed  CAS  Google Scholar 

  • Kaya S, Cetin ES, Aridogan BC, Arikan S, Demirci M (2007) Pathogenicity of Blastocystis hominis, a clinical reevaluation. Turk Parazitol Derg 31:184–187

    Google Scholar 

  • Kukoschke KG, Müller HE (1991) SDS-PAGE and immunological analysis of different axenic Blastocystis hominis strains. J Med Microbiol 35:35–39

    Article  PubMed  CAS  Google Scholar 

  • Lanuza MD, Carbajal JA, Villar J, Mir A, Borra’s R (1999) Soluble protein and antigenic heterogeneity in axenic Blastocystis hominis isolates: pathogenic implications. Parasitol Res 85:93–97

    Article  PubMed  CAS  Google Scholar 

  • Mansour NS, Mikhail EM, el Masry NA, Sabry AG, Mohareb EW (1995) Biochemical characterization of human isolates of Blastocystis hominis. J Med Microbiol 42:304–307

    Article  PubMed  CAS  Google Scholar 

  • Moe KT, Singh M, Ho LC, Tan SW, Chen XQ, Ng GC, Yap EH (1997) Experimental Blastocystis hominis infection in laboratory mice. Parasitol Res 83:319–325

    Article  PubMed  CAS  Google Scholar 

  • Müller HE (1994) Four serologically different groups within the species Blastocystis hominis. Zentralbl Bakteriol 280:403–408

    PubMed  Google Scholar 

  • Noël C, Dufernez F, Gerbod D, Edgcomb VP, Delgado-Viscogliosi P, Ho LC, Singh M, Wintjens R, Sogin ML, Capron M, Pierce R, Zenner L, Viscogliosi E (2005) Molecular phylogenies of Blastocystis isolates from different hosts: implications for genetic diversity, identification of species, and zoonosis. J Clin Microbiol 43:348–355

    Article  PubMed  Google Scholar 

  • Phillips BP, Zierdt CH (1976) Blastocystis hominis: pathogenic potential in human patients and in gnotobiotes. Exp Parasitol 39:358–364

    Article  PubMed  CAS  Google Scholar 

  • Rossignol JF, Kabil SM, Said M, Samir H, Younis AM (2005) Effect of nitazoxanide in persistent diarrhea and enteritis associated with Blastocystis hominis. Clin Gastroenterol Hepatol 3:987–991

    Article  PubMed  CAS  Google Scholar 

  • Santos HJ, Rivera WL (2009) Kinetic analysis of antibody responses to Blastocystis hominis in sera and intestinal secretions of orally infected mice. Parasitol Res 105(5):1303–1310

    Article  PubMed  Google Scholar 

  • Souppart L, Moussa H, Cian A, Sanciu G, Poirier P, El Alaoui H, Delbac F, Boorom K et al (2009) Subtype analysis of Blastocystis isolates from symptomatic patients in Egypt. Parasitol Res 106(2):505–511

    Article  PubMed  Google Scholar 

  • Svenungsson B, Lagergren A, Ekwall E, Evengard B, Hedlund KO, Karnell A, Lofdahl S, Svensson L, Weintraub A (2000) Enteropathogens in adult patients with diarrhea and healthy control subjects: a 1-year prospective study in a Swedish clinic for infectious diseases. Clin Infect Di 30:770–778

    Article  CAS  Google Scholar 

  • Tan TC, Suresh KG (2006) Predominance of amoeboid forms of Blastocystis hominis in isolates from symptomatic patients. Parasitol Res 98:189–193

    Article  PubMed  CAS  Google Scholar 

  • Tan SW, Ho LC, Moe KT, Chen XQ, Ng GC, Yap EH, Singh M (1996) Production and characterization of murine monoclonal antibodies to Blastocystis hominis. Int J Parasitol 26:375–381

    Article  PubMed  CAS  Google Scholar 

  • Tan SW, Singh M, Ho LC, Howe J, Moe KT, Chen XQ, Ng GC, Yap EH (1997) Survival of Blastocystis hominis clones after exposure to a cytotoxic monoclonal antibody. Int J Parasitol 27:947–954

    Article  PubMed  CAS  Google Scholar 

  • Tan KS, Ng GC, Quek E, Howe J, Ramachandran NP, Yap EH, Singh M (2000) Blastocystis hominis: a simplified, high-efficiency method for clonal growth on solid agar. Exp Parasitol 96:9–15

    Article  PubMed  CAS  Google Scholar 

  • Tan SW, Ibrahim M, Ng GC, Nasirudeen AMA, Ho LC, Yap EH, Singh M (2001) Exposure of Blastocystis species to a cytotoxic monoclonal antibody. Parasitol Res 87:534–538

    Article  PubMed  CAS  Google Scholar 

  • Tan SW, Yap EH, Singh M (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32:789–804

    Article  PubMed  CAS  Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Nat Acad Sci USA 76:4350–4354

    Article  PubMed  CAS  Google Scholar 

  • Upcroft JA, Dunn LA, Dommett LS, Healey A, Upcroft P, Boreham PFL (1989) Chromosomes of Blastocystis hominis. Int J Parasitol 19:879–883

    Article  PubMed  CAS  Google Scholar 

  • Zierdt CH (1988) Blastocystis hominis, a long misunderstood intestinal parasite. Parasitol Today 4:15–17

    Article  PubMed  CAS  Google Scholar 

  • Zierdt CH, Tan HK (1976) Ultrastructure and light microscope appearance of Blastocystis hominis in a patient with enteric disease. Z Parasitenkd 50:277–283

    Article  PubMed  CAS  Google Scholar 

  • Zierdt CH, Nagy B (1993) Antibody response to Blastocystis hominis infections. Ann Intern Med 118:985–986

    PubMed  CAS  Google Scholar 

  • Zierdt CH, Zierdt WS, Nagy B (1995) Enzyme-linked immunosorbent assay for detection of serum antibody to Blastocystis hominis in symptomatic infections. J Parasitol 81:127–129

    Article  PubMed  CAS  Google Scholar 

Download references

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Correspondence to Hala S. Elwakil.

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Gamra, M.M.A., Elwakil, H.S., El Deeb, H.K. et al. The potential use of 29 kDa protein as a marker of pathogenicity and diagnosis of symptomatic infections with Blastocystis hominis . Parasitol Res 108, 1139–1146 (2011). https://doi.org/10.1007/s00436-010-2156-8

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