Molecular Phylodiagnosis of Enterocytozoon bieneusi and Encephalitozoon intestinalis in Children with Cancer: Microsporidia in Malignancies as an Emerging Opportunistic Infection



Microsporidia may cause infection in both immunocompromised and immunocompetent populations. The best strategy to control microsporidiosis is obtaining thorough knowledge of its outbreak and pathogenicity.


Because of the lack of precise estimation of microsporidia prevalence among Iranian children with cancer, the current study aimed at evaluating the rate of intestinal microsporidia in children undergoing chemotherapy.


 Patients with cancer undergoing chemotherapy in a children’s hospital in Northwestern Iran were studied; 132 stool samples were collected and stained by the Weber and Ryan-blue modified trichrome staining techniques. The extracted DNA samples were evaluated by the nested polymerase chain reaction (PCR) method. All positive isolates were sequenced for genotyping and phylogenetic analysis.


A total of 17 (12.8%) samples were microscopically positive for microsporidia infection, whereas only 14 (10.6%) cases were positive based on nested PCR results. In the positive samples detected with nested PCR, the frequency of Enterocytozoon bieneusi and Encephalitozoon intestinalis infections was 71.4% (n = 10) and 28.6% (n = 4), respectively. After sequencing and phylogenetic analysis, the genotype of E. bieneusi was type D and the sequences of the isolated species were similar to those of the registered ones.


E. bieneusi is a major contributor to microsporidiosis in young immunocompromised patients in Iran. Microsporidia species are well-detected when confirmatory techniques such as molecular methods are in agreement with staining. So, to ensure this, a suggestion has been made to introduce a certain diagnostic test for microsporidiosis.

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This study was financially supported by Pediatric Health Research Center, Tabriz University of Medical Sciences, Iran. This is a report of a database from the master’s thesis of the first author registered in Tabriz University of Medical Sciences (Thesis number: 93/2–4/12).

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Correspondence to Mahmoud Mahami-Oskouei.

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Ghoyounchi, R., Mahami-Oskouei, M., Rezamand, A. et al. Molecular Phylodiagnosis of Enterocytozoon bieneusi and Encephalitozoon intestinalis in Children with Cancer: Microsporidia in Malignancies as an Emerging Opportunistic Infection. Acta Parasit. 64, 103–111 (2019).

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  • Enterocytozoon bieneusi
  • Encephalitozoon intestinalis
  • Children
  • Cancer
  • PCR
  • Iran