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. 2001 Jun;39(6):2248-53.
doi: 10.1128/JCM.39.6.2248-2253.2001.

Genotyping Encephalitozoon cuniculi by multilocus analyses of genes with repetitive sequences

L Xiao  1 , L Li, G S Visvesvara, H Moura, E S Didier, A A Lal
Affiliations

Genotyping Encephalitozoon cuniculi by multilocus analyses of genes with repetitive sequences

L Xiao et al. J Clin Microbiol. 2001 Jun.

Abstract

Encephalitozoon cuniculi infects a wide range of mammalian hosts. Three genotypes based on the number of GTTT repeats in the internal transcribed spacer (ITS) of the rRNA have been described, of which genotypes I and III have been identified in humans. In this study, the genetic diversity of E. cuniculi was examined at the polar tube protein (PTP) and spore wall protein I (SWP-1) loci. Nucleotide sequence analysis of the PTP gene divided 11 E. cuniculi isolates into three genotypes in congruence with the result of analysis of the ITS of the rRNA gene. The three PTP genotypes differed from one another by the copy number of the 78-bp central repeat as well as point mutations. These E. cuniculi isolates also differed from one another in the number of 15- and 36-bp repeats in the SWP-1 gene. In addition, some E. cuniculi isolates had heterogeneous copies of the SWP-1 gene with various numbers of repeats. Intragenotypic variation was also observed at the SWP-1 locus. Based on the length polymorphism and sequence diversities of the PTP and SWP-1 genes, two simple PCR tests were developed to differentiate E. cuniculi in clinical samples.

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Figures

FIG. 1
FIG. 1
Sequence diversity among three genotypes (I, II, and III) of E. cuniculi in the PTP gene. Dots denote sequence identity to genotype I and dashes denote nucleotide deletions. The repeat region is underlined.
FIG. 2
FIG. 2
Genotyping E. cuniculi isolates by PCR analysis of the PTP gene. (a) Differentiation of genotype III from genotypes I and II by eletrophoresis of PCR products: lanes 1 and 10, 100-bp ladders; lanes 2 (strain I), 4 (CDC:V385), 6 (CDC:V446), and 9 (CDC:V428A), genotype I; lane 8 (strain II), genotype II; and lanes 3 (CDC:V282), 5 (USP A-1), and 7 (3275), genotype III. (b) Differentiation of genotype I from genotype II by restriction digestion of PCR products with Sau96I: lanes 2 (strain I), 3 (CDC:V385), 4 (CDC:V446), and 6 (CDC:V428A), genotype I; and lane 5 (strain II), genotype II.
FIG. 3
FIG. 3
Genotyping E. cuniculi isolates by PCR analysis of SWP-1. Lane 1, 100-bp ladder; lanes 2 (CDC:V282), 4 (USP A-1), and 7 (USP A-2), genotype IIIa; lanes 3 (CDC:V428A), 6 (strain I), and 8 (CDC:V385), genotype Ia; lanes 5 (CDC:V449A) and 10 (strain III), genotype IIIb; lane 9 (CDC:V446), genotype Ib; lane 11 (strain II), genotype II.
FIG. 4
FIG. 4
Genetic relatedness of spore wall protein gene sequences based on a neighbor-joining analysis.
See this image and copyright information in PMC

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