Skip to main content
SpringerLink
Log in

PCR fingerprinting of Blastocystis isolated from symptomatic and asymptomatic human hosts

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Genomic DNA from 16 Blastocystis hominis isolates comprising of eight asymptomatic isolates (A1–A8) and eight symptomatic isolates (S1–S8) was amplified by arbitrarily primed polymerase chain reaction (AP-PCR) using 38 arbitrary 10-mer primers. Six primers (A10, B5, C20, D1, F6, and F10) generated reproducible DNA fingerprints. AP-PCR amplification revealed similar DNA fingerprints among all symptomatic isolates (S1–S8) with common bands at 850 bp using primer A10, 920 bp using primer B5, and 1.3 kbp using primer D1. Isolates A1, A3, A4, A5, A6, and A7 showed similar DNA banding patterns and all asymptomatic isolates (A1–A8) shared a major band at 1 kbp using primer B5. Isolates A2 and A8 showed distinct DNA banding patterns that differed from the remainder of the isolates. The results of the phylogenetic analyses showed that all symptomatic isolates (S1–S8) formed a clade with >70% similarity among the isolates and which were clearly separate from asymptomatic isolates A1, A3, A4, A5, A6, and A7. Asymptomatic isolates A2 and A8 formed two distinct and separate clades. AP-PCR revealed higher genetic variability within the asymptomatic isolates than within the symptomatic isolates. The present study suggests that AP-PCR can be a valuable method for differentiating between isolates of B. hominis and our results support the hypothesis that our asymptomatic and symptomatic B. hominis isolates may represent two different strains/species with varying pathogenic potential.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Arisue N, Hashimoto T, Yoshikawa H (2003) Sequence heterogeneity of the small subunit RNA genes among Blastocystis isolates. Parasitology 126:1–9

    Article  PubMed  CAS  Google Scholar 

  • Böhm-Gloning B, Knobloch J, Walderich B (1997) Five subgroups of Blastocystis hominis isolates from symptomatic and asymptomatic patients revealed by restriction site analysis of PCR amplified 16S-like rDNA. Trop Med Int Health 2:771–778

    Article  PubMed  Google Scholar 

  • Carbajal JA, Del Castillo L, Lanuza MD, Villar J, Borrás R (1997) Karyotypic diversity among Blastocystis hominis isolates. Int J Parasitol 27:941–945

    Article  PubMed  CAS  Google Scholar 

  • Clark CG (1997) Extensive genetic diversity in Blastocystis hominis. Mol Biochem Parasitol 87:79–83

    Article  PubMed  CAS  Google Scholar 

  • Ellsworth DL, Rittenhouse KD, Honeycutt RL (1993) Artifactual variation in randomly amplified polymorphic DNA banding patterns. Biotechniques 14:214–217

    PubMed  CAS  Google Scholar 

  • Fraga J, Rojas L, Sariego I, Sarría CA (2002) Optimization of random amplified polymorphic DNA techniques for its use in genetic studies of Trichomonas vaginalis isolates. Infect Genet Evol 2:73–75

    Article  PubMed  CAS  Google Scholar 

  • Guo ZG, Gross U, Johnson AM (1997) Toxoplasma gondii virulence markers identified by random amplified polymorphic DNA polymerase chain reaction. Parasitol Res 83:458–463

    Article  PubMed  CAS  Google Scholar 

  • Ho LC, Singh M, Suresh K, Ng GC, Yap EH (1994) A study of the karyotypic patterns of Blastocystis hominis by pulsed-field gradient electrophoresis. Parasitol Res 80:620–622

    Article  PubMed  CAS  Google Scholar 

  • Ho LC, Jeyaseelan K, Singh M (2001) Use of elongation-1 alpha gene in a polymerase chain reaction-based restriction-fragment-length polymorphism analysis of genetic heterogeneity among Blastocystis species. Mol Biochem Parasitol 112:287–291

    Article  PubMed  CAS  Google Scholar 

  • Hoevers J, Holman P, Logan K, Hommel M, Ashford R, Snowden K (2000) Restriction-fragment-length polymorphism analysis of small subunit rRNA genes of Blastocystis hominis isolates from geographically diverse human hosts. Parasitol Res 86:57–61

    Article  PubMed  CAS  Google Scholar 

  • Init I, Mak JW, Lokman Hakim S, Yong HS (1999) Strain differences in Blastocystis isolates as detected by a single set of polymerase chain reaction primers. Parasitol Res 85:131–134

    Article  PubMed  CAS  Google Scholar 

  • Jones WR (1946) The experimental infection of rats with Entamoeba histolytica. Ann Trop Med Parasitol 40:130

    Google Scholar 

  • Lanuza MD, Carbajal JA, Villar J, Borrás R (1997) Description of an improved method for Blastocystis hominis culture and axenization. Parasitol Res 83:60–63

    Article  PubMed  CAS  Google Scholar 

  • Mackenstedt U, Johnson AM (1995) Genetic differentiation of pathogenic and non-pathogenic strains of Entamoeba histolytica by random amplified polymorphic DNA polymerase chain reaction. Parasitol Res 81:217–221

    Article  PubMed  CAS  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 

  • Paintlia AS, Mahajan RC, Chakraborti A, Sehgel R, Ganguly NK (1999) Characterization of Giardia lamblia groups A and B from North India by isoenzyme and random amplified polymorphic DNA analysis. Parasitol Res 85:510–512

    Article  PubMed  CAS  Google Scholar 

  • Rajah S, Suresh K, Vennila GD, Khairual Anuar A, Saminathan R (1997) Small forms of Blastocystis hominis. Int Med Res J 1:93–96

    Google Scholar 

  • Snowden K, Logan K, Blozinski C, Hoevers J, Holman P (2000) Restriction-fragment-length polymorphism analysis of small-subunit rRNA genes of Blastocystis isolates from animal hosts. Parasitol Res 86:62–66

    Article  PubMed  CAS  Google Scholar 

  • Stenzel DJ, Boreham PFL (1996) Blastocystis hominis revisited. Clin Microbiol Rev 9:563–584

    PubMed  CAS  Google Scholar 

  • Suresh K, Howe J, Chong SY, Ng GC, Ho LC, Loh AK, Ramachandran NP, Yap EH, Singh M (1994) Ultrastructural changes during in vitro encystment of Blastocystis hominis. Parasitol Res 80:327–335

    Article  PubMed  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 KSW, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spot in terra incognita. Int J Parasitol 32:789–804

    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 

  • Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218

    Article  PubMed  CAS  Google Scholar 

  • Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic acids Res 18:6531–6535

    Article  PubMed  CAS  Google Scholar 

  • Yoshikawa H, Nagono I, Yap EH, Singh M, Takahashi Y (1996) DNA polymorphism revealed by arbitrary primers polymerase chain reaction among Blastocystis strains isolated from humans, a chicken and a reptile. J Eukaryot Microbiol 43:127–130

    Article  PubMed  CAS  Google Scholar 

  • Yoshikawa H, Abe N, Iwasawa M, Kitano S, Nagano I, Wu Z, Takahashi Y (2000) Genomic analysis of Blastocystis hominis strains isolated from two long-term health care facilities. J Clin Microbiol 38:1324–1330

    PubMed  CAS  Google Scholar 

  • Yoshikawa H, Wu Z, Kimata I, Iseki M, Ali IK, Hossain MB, Zaman V, Haque R, Takahashi Y (2004) Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 93:22–29

    Google Scholar 

  • Zaman V (1997) Phase-contrast microscopy of cell division in Blastocystis hominis. Ann Trop Med Parasitol 91:223–224

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This study was funded by IRPA grant R&D 06-02-03-1012 from the Malaysian Government. We thank the Department of Veterinary Parasitology, Murdoch University, Perth, Australia, the Department of Parasitology, Aga Khan University, Pakistan, and the Department of Parasitology, Mahidol University, Thailand for providing Blastocystis samples.

Author information

Authors and Affiliations

  1. Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia

    T. C. Tan & K. G. Suresh

  2. Institute for Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia

    K. L. Thong

  3. Scottish Parasite Diagnostic Laboratory, Stobhill Hospital, Glasgow G21 3UW, Scotland, UK

    H. V. Smith

Authors
  1. T. C. Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. G. Suresh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. L. Thong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. V. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K. G. Suresh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tan, T.C., Suresh, K.G., Thong, K.L. et al. PCR fingerprinting of Blastocystis isolated from symptomatic and asymptomatic human hosts. Parasitol Res 99, 459–465 (2006). https://doi.org/10.1007/s00436-006-0177-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-006-0177-0

Keywords

Search

Navigation