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

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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.

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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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  1. Ibne Karim M. D. Ali

    Present address: Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London, WC1E 7HT, UK

Authors and Affiliations

  1. Department of Biological Science, Faculty of Science, Nara Women’s University, Kitauoya-Nishimachi, Nara 630–8506, Japan

    Hisao Yoshikawa

  2. Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500–8705, Japan

    Zhiliang Wu & Yuzo Takahashi

  3. Department of Medical Zoology, Osaka City University Graduate School of Medicine, Asahimachi, Abeno, Osaka 545–8585, Japan

    Isao Kimata

  4. Department of Parasitology, Graduate School of Medical Science, Kanazawa University, Takaramachi, Kanazawa 920–8640, Japan

    Motohiro Iseki

  5. International Centre for Diarrhoeal Diseases Research, G.P.O. Box 128, 1000, Dhaka, Bangladesh

    Ibne Karim M. D. Ali, Momammad B. Hossain & Rashidul Haque

  6. Department of Microbiology, Aga Khan University, Stadium Road, P.O. Box 3500, 74800, Karachi, Pakistan

    Viqar Zaman

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  1. Hisao Yoshikawa
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Correspondence to Hisao Yoshikawa.

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Yoshikawa, H., Wu, Z., Kimata, I. et al. Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 92, 22–29 (2004). https://doi.org/10.1007/s00436-003-0995-2

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  • DOI: https://doi.org/10.1007/s00436-003-0995-2

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