Parasitology Research

, Volume 92, Issue 1, pp 22–29 | Cite as

Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries

  • Hisao Yoshikawa
  • Zhiliang Wu
  • Isao Kimata
  • Motohiro Iseki
  • Ibne Karim M. D. Ali
  • Momammad B. Hossain
  • Viqar Zaman
  • Rashidul Haque
  • Yuzo Takahashi
Original Paper

Abstract

Since the genotype of human Blastocystis hominis isolates is highly polymorphic, PCR-based genotype classification using known sequenced-tagged site (STS) primers would allow the identification or classification of different genotypes. Five populations of human B. hominis isolates obtained from Japan, Pakistan, Bangladesh, Germany, and Thailand were subjected to genotype analysis by using seven kinds of STS primers. Ninety-nine out of 102 isolates were identified as one of the known genotypes, while one isolate from Thailand showed two distinct genotypes and two isolates from Japan were negative with all the STS primers. The most dominant genotype among four populations, except for all four isolates from Thailand, was subtype 3 and it varied from 41.7% to 92.3%. The second most common genotype among four populations was either subtype 1 (7.7–25.0%) or subtype 4 (10.0–22.9%). Subtype 2, subtype 5, and/or subtype 7 were only rarely detected among the isolates from Japan and Germany, while subtype 6 was not detected. The phylogenetic position of the two isolates which were negative with all STS primers, was inferred from the small subunit rRNA (SSU rRNA) genes with the known sequence data of 20 Blastocystis isolates. Since the two isolates were positioned in an additional clade in the phylogenetic tree, this suggested they were a new genotype. These results demonstrated that PCR-based genotype classification is a powerful tool with which to analyse genotypes of Blastocystis isolates obtained from clinical samples. In addition, two groups of the isolates from 15 symptomatic and 11 asymptomatic patients in Bangladesh were compared with the PCR-based subtype classification. Since both groups were only classified into two distinct genotypes of subtype 1 or subtype 3 and no statistically significant difference was observed between the two groups, in this study it could not be shown that the specific genotype correlated with the pathogenic potential of B. hominis.

Notes

Acknowledgements

We express our gratitude to Dr S. Tesana, Khon Kaen University, Thailand and Dr H. M. Seiz, Bonn University, Germany for collecting the faecal samples in Thailand and Germany, respectively, and thank Dr T. Hashimoto, University of Tsukuba for the phylogenetic analysis. This work was supported by a grant from the Japan Society for the Promotion of Science to H. Y. (C-13670245) and by a grant from ICDDR, B Centre for Health and Population Research with the support of the University of Virginia (NIH grant, AI-43596) to A. I.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Hisao Yoshikawa
    • 1
  • Zhiliang Wu
    • 2
  • Isao Kimata
    • 3
  • Motohiro Iseki
    • 4
  • Ibne Karim M. D. Ali
    • 5
    • 7
  • Momammad B. Hossain
    • 5
  • Viqar Zaman
    • 6
  • Rashidul Haque
    • 5
  • Yuzo Takahashi
    • 2
  1. 1.Department of Biological Science, Faculty of ScienceNara Women’s UniversityKitauoya-NishimachiJapan
  2. 2.Department of ParasitologyGifu University School of MedicineJapan
  3. 3.Department of Medical ZoologyOsaka City University Graduate School of MedicineAbeno, Osaka 545–8585Japan
  4. 4.Department of Parasitology, Graduate School of Medical ScienceKanazawa UniversityTakaramachi, Kanazawa 920–8640Japan
  5. 5.International Centre for Diarrhoeal Diseases ResearchDhakaBangladesh
  6. 6.Department of MicrobiologyAga Khan UniversityKarachiPakistan
  7. 7.Department of Infectious and Tropical DiseasesLondon School of Hygiene and Tropical MedicineLondonUK

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