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Journal of Clinical Microbiology, September 2003, p. 4166-4171, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4166-4171.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Intraspecies Genotype Variability of the Microsporidian Parasite Encephalitozoon hellem

María Haro, Carmen del Águila,* Soledad Fenoy, and Nuno Henriques-Gil

Departamento de Biología Celular, Bioquímica y Biología Molecular, Facultad de Ciencias Experimentales y de la Salud, Universidad San Pablo-CEU, 28668 Boadilla del Monte, Madrid, Spain

Received 20 November 2002/ Returned for modification 23 January 2003/ Accepted 24 June 2003

Seven isolates of Encephalitozoon hellem from human immunodeficiency virus-positive patients were genotyped through a series of markers: the internal transcribed spacer (ITS) of ribosomal DNA, the polar tube protein (PTP) gene, and two intergenic spacers (IGS-TH and IGS-HZ) whose polymorphism is newly reported. The genome markers were all analyzed at three levels: PCR amplification followed by polyacrylamide gel electrophoresis, single-strand conformation analysis (SSCA), and DNA sequencing. The polymorphisms detected involve insertions/deletions and point mutations. SSCA can distinguish any pair of sequences, even those differing by a single base pair. The different isolates studied fit into the previously described ITS genotype 1A, except one which seems to be a 2A derivative variant (2D). When PTP and the new markers IGS-TH and IGS-HZ were analyzed, most of the isolates displayed different genotypes, demonstrating that E. hellem has a strong intraspecies variability. A set of markers such as those used here may be very useful in genotyping of clinical samples and in the assessment of epidemiological relationships among E. hellem strains.

* Corresponding author. Mailing address: Facultad de Ciencias Experimentales y de la Salud, Urbanización Montepríncipe, 28668 Boadilla del Monte, Madrid, Spain. Phone: 34-91-372-47-96/84. Fax: 34-91-351-04-96. E-mail: cagupue{at}ceu.es.

Journal of Clinical Microbiology, September 2003, p. 4166-4171, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4166-4171.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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