Detection and Differentiation between Enterocytozoon bieneusi and Encephalitozoon intestinalis Species in Cancer Patient's Stools Using PCR Compared with Different Staining Methods

Background: Microsporidia are obligate intracellular parasites that are recognized as important opportunistic pathogens of immunocompromised patients especially HIV/AIDS and cancer patients. Enterocytozoon (Ent) bieneusi and Encephalitozoon (Enc) intestinalis are the most prevalent species in humans causing intestinal microsporidiosis. Objective: To evaluate the efficacy of regular PCR in the detection of and differentiation between Ent. bieneusi and Enc. intestinalis in the stool samples of cancer patients in comparison to microscopic staining techniques. Methodology: 100 cancer patients suffering from leukemia (28), cancer lung (10), cancer liver (24), cancer colon (16) and cancer breast (22) were included in this study. They were divided into diarrheic and nondiarrheic groups. The stool samples were concentrated, preserved and examined using modified trichrome blue stain (MTB), acridine orange (AO) and calcofluor white (CFW) in addition to regular PCR. Results: PCR technique showed the highest sensitivity, while microscopic examination revealed that CFW was more sensitive than other stains. The infection rate was significantly higher among diarrheic than nondiarrheic cancer patients by all used methods. Enc. intestinalis was the only species detected in positive stool samples except for one case that did not amplify with either Enc. intestinalis or Ent. bieneusi specific primers. Percentage detection of microsporidial infection using PCR was higher among leukemic patients than other groups of cancer patients. Conclusion: Regular PCR can be used as a confirmatory test beside MTB and CFW as screening tests for microsporidian species differentiation and proper clinical management and therapy.

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