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

Parasitology Research - Tập 92 - Trang 22-29 - 2003
Hisao Yoshikawa1, Zhiliang Wu2, Isao Kimata3, Motohiro Iseki4, Ibne Karim M. D. Ali5,6, Momammad B. Hossain5, Viqar Zaman7, Rashidul Haque5, Yuzo Takahashi2
1Department of Biological Science, Faculty of Science, Nara Women’s University, Kitauoya-Nishimachi, Japan
2Department of Parasitology, Gifu University School of Medicine, Japan
3Department of Medical Zoology, Osaka City University Graduate School of Medicine, Abeno, Osaka 545–8585, Japan
4Department of Parasitology, Graduate School of Medical Science, Kanazawa University, Takaramachi, Kanazawa 920–8640, Japan
5International Centre for Diarrhoeal Diseases Research, Dhaka, Bangladesh
6Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, UK
7Department of Microbiology, Aga Khan University, Karachi, Pakistan

Tóm tắt

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.

Tài liệu tham khảo

Abe N, Wu Z, Yoshikawa H (2003a) Molecular characterization of Blastocystis isolates from birds by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene. Parasitol Res 89:393–396

Abe N, Wu Z, Yoshikawa H (2003b) Molecular characterization of Blastocystis isolates from primates. Vet Parasitol 113:321–325

Abe N, Wu Z, Yoshikawa H (2003c) Zoonotic genotypes of Blastocystis hominis detected in cattle and pigs by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene. Parasitol Res 90:124–128

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

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

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

Chen XQ, Singh M, Ho LC, Tan SW, Ng GC, Moe KT, Yap EH (1997) Description of a Blastocystis species from Rattus norvegicus. Parasitol Res 83:313–318

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

Galantowicz BB, Illueca MD, Levy J, Rayburn JL, Weinstock DJ (1993) Neonatal Blastocystis hominis diarrhea. Pediatr Infect Dis 12:345–347

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

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

Hussain R, Jaferi W, Zuberi S, Baqai R, Abrar N, Ahmed A, Zaman V (1997) Significantly increased IgG2 subclass antibody levels to Blastocystis hominis in patients with irritable bowel syndrome. Am J Trop Med Hyg 56:301–306

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

Kaneda Y, Horiki N, Cheng XJ, Fujita Y, Maruyama M, Tachibana H (2001) Ribodemes of Blastocystis hominis isolated in Japan. Am J Trop Med Hyg 65:393–396

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

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

Leelayoova S, Taamasri P, Rangsin R, Naaglor T, Thathaisong U, Mungthin M (2002) In-vitro cultivation: a sensitive method for detecting Blastocystis hominis. Ann Trop Med Parasitol 96:803–807

Nimri LF (1993) Evidence of an epidemic of Blastocystis hominis infections in preschool children in northern Jordan. J Clin Microbiol 31:2706–2708

Noel C, Peyronnet C, Gerbod D, Edgcomb VP, Delgado-Viscogliosi P, Sogin ML, Capron M, Viscogliosi E, Zenner L (2003) Phylogenetic analysis of Blastocystis isolates from different hosts based on the comparison of small-subunit rRNA gene sequences. Mol Biochem Parasitol 126:119–123

Salim HR, Kumar GS, Vellayan S, Mak JW, Anuar AK, Init I, Vennila GD, Saminathan R, Ramakrishnan K (1999) Blastocystis in animal handlers. Parasitol Res 85:1032–1033

Schonian G, Akuffo H, Lewin S, Maasho K, Nylen S, Pratlong F, Eisenberger CL, Schnur LF, Presber W (2000) Genetic variability within the species Leishmania aethiopica does not correlate with clinical variations of cutaneous leishmaniasis. Mol Biochem Parasitol 106:239–248

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

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

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

Windsor JJ, Macfarlane L, Hughes-Thapa G, Jones SKA, Whiteside TM (2002) Incidence of Blastocystis hominis in faecal samples submitted for routine microbiological analysis. Br J Biomed Sci 59:154–157

Yoshikawa H, Nagano 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 Euk Microbiol 43:127–130

Yoshikawa H, Nagano I, Wu Z, Yap EH, Singh M, Takahashi Y (1998) Genomic polymorphism among Blastocystis strains and development of subtype-specific diagnostic primers. Mol Cell Probes 12:153–159

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

Yoshikawa H, Wu Z, Nagano I, Takahashi Y (2003) Molecular comparative studies among Blastocystis isolates obtained from humans and animals. J Parasitol 89:585–594

Zaman V, Khan KZ (1994) A comparison of direct microscopy with culture for the diagnosis of Blastocystis hominis. Southeast Asian J Trop Med Public Health 25:792–793

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

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Parasitology Research

Tập 92

22-29

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