Molecular Phylodiagnosis of Enterocytozoon bieneusi and Encephalitozoon intestinalis in Children with Cancer: Microsporidia in Malignancies as an Emerging Opportunistic Infection
- 34 Downloads
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.
KeywordsEnterocytozoon bieneusi Encephalitozoon intestinalis Children Cancer PCR Iran
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).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- 2.Askari Z., Mirjalali H., Mohebali M., Zarei Z., Shojaei S., Rezaeian T., Rezaeian M. 2015. Molecular detection and identification of zoonotic microsporidia spore in fecal samples of some animals with close-contact to human. Iranian Journal of Parasitology, 10, 381–388Google Scholar
- 3.Berahmat R., Mahami-Oskouei M., Rezamand A., Spotin A., Aminisani N., Ghoyounchi R., Madadi S. 2017. Cryptosporidium infection in children with cancer undergoing chemotherapy: how important is the prevention of opportunistic parasitic infections in patients with malignancies? Parasitology Research, 116, 2507–2515. https://doi.org/10.1007/s00436-017-5560-5 CrossRefGoogle Scholar
- 4.Cali A., Takvorian N.R.P.M. 2011. Microsporidiosis, p 61–76. In Meyers WM, Firpo A, Wear DJ (ed), Topics on the pathology of protozoan and invasive arthropod diseases. Armed Forces Institute of Pathology (AFIP), Washington, DC. http://oai.dtic.mil/oai/oai?verb_getRecord&metadataPrefix_html&identifier_ADA545141.
- 5.Chandramathi S., Suresh K., Anita Z.B., Kuppusamy U.R. 2012. Infections of Blastocystis hominis and microsporidia in cancer patients: are they opportunistic? Transactions of the Royal Society of Tropical Medicine and Hygiene, 106, 267–269. https://doi.org/10.1016/j.trstmh.2011.12.008 CrossRefGoogle Scholar
- 7.Desportes I., Charpentier YL., Galian A., Bernard F., Cochand‐Priollet B., Lavergne A., Ravisse P., Modigliani R. 1985. Occurrence of a new microsporidan: Enterocytozoon bieneusi ng, n. sp., in the enterocytes of a human patient with AIDS. The Journal of Protozoology, 32, 250–254.CrossRefGoogle Scholar
- 8.Didier E.S., Vossbrinck C.R., Stovall M.E., Green L.C., Bowers L., Fredenburg A., Didier P.J. 2004. Diagnosis and epidemiology of microsporidian infections in humans. The Southeast Asian Journal of Tropical Medicine and Public Health, 35, 65–81.Google Scholar
- 9.Didier E.S., Weiss L.M. 2006. Microsporidiosis: current status. Current Opinion in Infectious Diseases, 19, 485–492. https://doi.org/10.1097/01.qco.0000244055.46382.23 CrossRefGoogle Scholar
- 10.El Sobky M., El Nahas N. 2012. Detection and differentiation between Enterocytozoon bieneusi and Encephalitozoon intestinalis species in cancer patient’s stools using PCR compared with different staining methods. Parasitology United Journal, 5, 20–26.Google Scholar
- 11.Eligio-García L., Cano-Estrada A., Cortés-Campos A., Medina-Sansón A., Jiménez-Cardoso E. 2013. Identification of Microsporidium spp. in patients with acute lymphoblastic leukemia. Boletín Médico del Hospital Infantil de México, 70, 26–30.Google Scholar
- 12.Ghorbanzadeh B., Sadraei J., Emadi H. 2012. Diagnosis of Cryptosporidium and intestinal Microsporidia in HIV/AIDS patients with staining and PCR methods on 16srRNA gen. Arak Medical University Journal, 15, 37–47.Google Scholar
- 13.Ghoyounchi R., Ahmadpour E., Spotin A., Mahami-Oskouei M., Rezamand A., Aminisani N., Ghojazadeh M., Berahmat R., Mikaeili-Galeh T. 2017. Microsporidiosis in Iran: a systematic review and meta-analysis. Asian Pacific Journal of Tropical Medicine, 10, 341–350. https://doi.org/10.1016/j.apjtm.2017.03.017 CrossRefGoogle Scholar
- 22.Mirjalali H., Mirhendi H., Meamar A.R., Mohebali M., Askari Z., Mirsamadi E.S., Rezaeian M. 2015. Genotyping and molecular analysis of Enterocytozoon bieneusi isolated from immunocompromised patients in Iran. Infection, Genetics and Evolution, 36, 244–249. https://doi.org/10.1016/j.meegid.2015.09.022 CrossRefGoogle Scholar
- 23.Mirjalali H., Mohebali M., Mirhendi H., Gholami R., Keshavarz H., Meamar A.R., Rezaeian M. 2014. Emerging intestinal microsporidia infection in HIV +/AIDS patients in Iran: microscopic and molecular detection. Iranian Journal of Parasitology, 9, 149–154.Google Scholar
- 24.Nadham Kadham M., Maysloon A.A.S. 2012. Microsporidiosis among children with malignant diseases in Basrah, Iraq. Pakistan Journal of Medical Sciences, 28, 621–624.Google Scholar
- 25.Nasarudin S.N., Zainudin N.S., Bernadus M., Nawi A.M., Hanafiah A., Osman E. 2015. Loop-mediated isothermal amplification for rapid molecular detection of Enterocytozoon bieneusi in faecal specimens. Journal of Medical Microbiology, 64, 1329–1334. https://doi.org/10.1099/jmm.0.000156 CrossRefGoogle Scholar
- 28.Ryan N.J., Sutherland G., Coughlan K., Globan M., Doultree J., Marshall J., Baird R., Pedersen J., Dwyer B. 1993. A new trichrome-blue stain for detection of microsporidial species in urine, stool, and nasopharyngeal specimens. Journal of Clinical Microbiology, 31, 3264–3269.Google Scholar
- 29.Sak B., Brady D., Pelikánová M., Květoňová D., Rost M., Kostka M., Tolarová V., Hůzová Z., Kváč M. 2011. Unapparent microsporidial infection among immunocompetent humans in the Czech Republic. Journal of Clinical Microbiology, 49, 1064–1070. https://doi.org/10.1128/jcm.01147-10 CrossRefGoogle Scholar
- 32.Tamura K., Peterson D., Peterson N., Stecher G., Nei M., Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731–2739. https://doi.org/10.1093/molbev/msr121 CrossRefGoogle Scholar
- 33.Tavalla M., Mardani-Kateki M., Abdizadeh R., Nashibi R., Rafie A., Khademvatan S. 2017. Molecular identification of Enterocytozoon bieneusi and Encephalitozoon spp. in immunodeficient patients in Ahvaz, Southwest of Iran. Acta Tropica, 172, 107–112. https://doi.org/10.1016/j.actatropica.2017.04.015 CrossRefGoogle Scholar
- 35.Weber R., Bryan R.T., Owen R.L., Wilcox CM., Gorelkin L., Visvesvara G.S. 1992. Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. The enteric opportunistic infections working group. The New England Journal of Medicine, 326, 161–166. https://doi.org/10.1056/nejm199201163260304 CrossRefGoogle Scholar
- 36.Zainudin N.S., Nasarudin S.N.S., Periyasamy P., Moktar N., Noordin R., Osman E. 2016. Diagnosis of disseminated microsporidiosis: detection of circulating Enterocytozoon bieneusi DNA in blood of HIV/AIDS patients. Tropical Biomedicine, 33, 761–770.Google Scholar