Frequency of Microsporidial Infection in Immunocompromised Patients with Staining and Molecular Methods Based on Internal Transcribed Spacer Region Gene in Two Cities of Southwest Iran during 2013-2014


  • Ahvaz Jundishapur University of Medical Sciences, Department of Medical Parasitology and Mycology, Abadan Arvand International Division, Ahwaz, Iran, Islamic Republic of
  • Ahvaz Jundishapur University of Medical Sciences, Health Research Institute, Infectious and Tropical Diseases Research Center, Ahwaz, Iran, Islamic Republic of
  • Ahvaz Jundishapur University of Medical Sciences, Department of Medical Parasitology, Faculty of Medicine, Ahwaz, Iran, Islamic Republic of
  • Ahvaz Jundishapur University of Medical Sciences, Abadan Arvand International Division, Ahwaz, Iran, Islamic Republic of


Microsporidiosisis anopportunistic infection frequently diagnosed in immunocompromised patients. Microsporidiosis causes symptomatic or asymptomatic infections immunocompromised/immunosuppressed patients may experience severe and disseminated microsporidiosis. Fecal samples were taken from 186 patients with human immunodeficiency virus (86) and cancer (100). Microsporidia spp. were detected 27.9% (24/86) and 11.6% (10/86) in HIV + patients, 18% (18/100) and 7% (7/100) of cancer patients using polymerase chain reaction (PCR) and trichrome staining respectively. Enterocytozoon bieneusi was detected in 28(66.7%) patients, while Encephalitozoon intestinalis was detected in 10 (23.8%) patients. Encephalitozoon cuniculi was detected in only one patient, while Encephalitozoon hellem was detected in three patients. Microsporidia was significantly higher in patients with diarrhea than those without diarrhea (P = 0.001). CD4 T cells blood count was significantly lower (<200 cells/μl blood) in HIV+ patients with Microsporidia (P = 0.001). Also, there was significant difference between contact with animals with positive results of HIV+ and cancer patients (P< 0.005). This study confirmed that PCR technique was more sensitive than trichrome staining for diagnosis. Moreover, intestinal microsporidiosis caused diarrhea in HIV+ and cancer patients. This influenced treatment. Therefore, early diagnosis of microsporidiosis is effective on treatment strategies.


HIV, Immunocompromised Patients, Iran, Microsporidia, PCR

Subject Discipline

Pharmacy and Pharmacology

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Didier ES, Didier PJ, Snowden KF, Shadduck JA. Microsporidiosis in mammals. Microbes and Infection. 2000;2(6):709–20.

Didier ES, Weiss LM. Microsporidiosis: current status. Current opinion in infectious diseases. 2006;19(5):485.

3.Eligio-García L, Cano-Estrada A, Cortés-Campos A, Medina-Sansón A, Jiménez-Cardoso E. Identification of Microsporidium spp. in patients with acute lymphoblastic leukemia. Boletín Médico del Hospital Infantil de México. 2013;70(1):26–30.

Dascomb K, Frazer T, Clark RA, Kissinger P, Didier E. Microsporidiosis and HIV. JAIDS Journal of Acquired Immune Deficiency Syndromes. 2000;24(3):290–2.

Dworkin MS, Buskin SE, Davidson AJ, Cohn DL, Morse A, Inungu J, et al. Prevalence of intestinal microsporidiosis in human immunodeficiency virus-infected patients with diarrhea in major United States cities. Revista do Instituto de Medicina Tropical de São Paulo. 2007;49(6):339–42.

Galván A, Sánchez AM, Valentín MP, Henriques-Gil N, Izquierdo F, Fenoy S, et al. First cases of microsporidiosis in transplant recipients in Spain and review of the literature. Journal of Clinical Microbiology. 2011;49(4):1301–6.

Wichro E, Hoelzl D, Krause R, Bertha G, Reinthaler F, Wenisch C. Microsporidiosis in travel-associated chronic diarrhea in immune-competent patients. The American Journal of Tropical Medicine and Hygiene. 2005;73(2):285–7.

Sokolova OI, Demyanov AV, Bowers LC, Didier ES, Yakovlev AV, Skarlato SO, et al. Emerging microsporidian infections in Russian HIV-infected patients. Journal of Clinical Microbiology. 2011;49(6):2102–8.

9.Sak B, Brady D, Pelikánová M, Květoňová D, Rost M, Kostka M, et al. Unapparent microsporidial infection among immunocompetent humans in the Czech Republic. Journal of Clinical Microbiology. 2011;49(3):1064–70.

Franzen C. Microsporidia: a review of 150 years of research. Open Parasitol J. 2008;2:1–34.

WHO. Cancer control programe.NCD country profiles. 2011.

Didier ES. Microsporidiosis: an emerging and opportunistic infection in humans and animals. Acta Tropica. 2005;94(1):61–76.

Botero JH, Castaño A, Montoya MN, Ocampo NE, Hurtado MI, Lopera MM. A preliminary study of the prevalence of intestinal parasites in immunocompromised patients with and without gastrointestinal manifestations. Revista do Instituto de Medicina Tropical de São Paulo. 2003;45(4):197–200.

Karaman U, Atambay M, Daldal N, Colak C. The prevalence of Microsporidium among patients given a diagnosis of cancer. Turkiye parazitolDerg. 2008;32(2):109–12.

Mirjalali H, Mohebali M, Mirhendi H, Gholami R, Keshavarz H, Meamar AR, et al. Emerging intestinal microsporidia infection in HIV+/AIDS patients in Iran: microscopic and molecular detection. Iranian Journal of Parasitology. 2014;9(2):149–54.

Agholi M, Hatam GR, Motazedian MH. HIV/AIDS-associated opportunistic protozoal diarrhea. AIDS Research and Human Retroviruses. 2013;29(1):35–41.

Agholi M, Hatam GR, Motazedian MH. Microsporidia and coccidia as causes of persistence diarrhea among liver transplant children: incidence rate and species/genotypes. The Pediatric Infectious Disease Journal. 2013;32(2):185–7.

18.Ghorbanzadeh B, Sadraie J, Emadi Kuchak H. Diagnosis of Cryptosporidium and intestinal Microsporidia in HIV/ AIDS patients with staining and PCR methods on 16srRNA gen. Arak Medical University Journal. 2012;15(7):37–47.

Franzen C, Müller A. Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia. Clinical Microbiology Reviews. 1999;12(2):243–85.

Ghosh K, Weiss LM. Molecular diagnostic tests for microsporidia. Interdisciplinary Perspectives on Infectious Diseases. 2009; 2009(1);1–13.

Subrungruang I, Mungthin M, Chavalitshewinkoon-Petmitr P, Rangsin R, Naaglor T, Leelayoova S. Evaluation of DNA extraction and PCR methods for detection of Enterocytozoon bienuesi in stool specimens. Journal of Clinical Microbiology. 2004;42(8):3490–4.

Katzwinkel‐Wladarsch S, Lieb M, Heise W, Löscher T, Rinder H. Direct amplification and species determination of microsporidian DNA from stool specimens. Tropical Medicine and International Health. 1996;1(3):373–8.

Bendall R, Chiodini P. New diagnostic methods for parasitic infections. Current Opinion in Infectious Diseases. 1993;6(3):318–22.

Velásquez JN, Carnevale S, Guarnera EA, Labbé JH, Chertcoff A, Cabrera MG, et al. Detection of the microsporidian parasite Enterocytozoon bieneusi in specimens from patients with AIDS by PCR. Journal of Clinical Microbiology. 1996;34(12):3230–2.

Bryan R, Schwartz D. Epidemiology of microsporidiosis. The Microsporidia and Microsporidiosis. 1999; 1(1):502– 16.

Coyle CM, Wittner M, Kotler DP, Noyer C, Orenstein JM, Tanowitz HB, et al. Prevalence of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon (Septata) intestinalis among patients with AIDS-related diarrhea: determination by polymerase chain reaction to the microsporidian small-subunit rRNA gene. Clinical Infectious Diseases. 1996;23(5):1002–6.

Espern A, Morio F, Miegeville M, Illa H, Abdoulaye M, Meyssonnier V, et al. Molecular study of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon intestinalis among human immunodeficiency virus-infected patients from two geographical areas: Niamey, Niger, and Hanoi, Vietnam. Journal of Clinical Microbiology. 2007;45(9):2999–3002.

Breton J, Bart-Delabesse E, Biligui S, Carbone A, Seiller X, Okome-Nkoumou M, et al. New highly divergent rRNA sequence among biodiverse genotypes of Enterocytozoon bieneusi strains isolated from humans in Gabon and Cameroon. Journal of Clinical Microbiology. 2007;45(8):2580–9.

Endeshaw T, Kebede A, Verweij JJ, Zewide A, Tsige K, Abraham Y, et al. Intestinal microsporidiosis in diarrheal patients infected with human immunodeficiency virus-1 in Addis Ababa, Ethiopia. Japanese Journal of Infectious Diseases. 2006;59(5):306.

Ojuromi OT, Izquierdo F, Fenoy S, Fagbenro-Beyioku A, Oyibo W, Akanmu A. Identification and characterization of microsporidia from fecal samples of HIV-positive patients from Lagos, Nigeria. PloS One. 2012;7(4): e35239.

Brasil P, de Lima DB, de Paiva DD, de Castro Lobo MS, Sodré FC, da Silva SP, et al. Clinical and diagnostic aspects of intestinal microsporidiosis in HIV-infected patients with chronic diarrhea in Rio de Janeiro, Brazil. Revista do Instituto de Medicina Tropical de Sao Paulo. 2000;42(6):299– 304.

Kulkarni S, Kairon R, Sane S, Padmawar P, Kale V, Thakar M, et al. Opportunistic parasitic infections in HIV/AIDS patients presenting with diarrhoea by the level of immunesuppression. 2009.

Chacin-Bonilla L, Panunzio AP, Monsalve-Castillo FM, Parra-Cepeda IE, Martinez R. Microsporidiosis in Venezuela: Prevalence of intestinal microsporidiosis and its contribution to diarrhea in a group of Human Immunodeficiency Virus–infected patients from Zulia State. The American Journal of Tropical Medicine and Hygiene. 2006;74(3):482– 6.

Chabchoub N, Abdelmalek R, Mellouli F, Kanoun F, Thellier M, Bouratbine A, et al. Genetic identification of intestinal microsporidia species in immunocompromised patients in Tunisia. The American Journal of Tropical Medicine and Hygiene. 2009;80(1):24–7.

Saigal K, Sharma A, Sehgal R, Sharma P, Malla N, Khurana S. Intestinal microsporidiosis in India: A two year study. Parasitology International. 2013;62(1):53–6.

Sarfati C, Bourgeois A, Menotti J, Liégeois F, Moyou-Somo R, Delaporte E, et al. Prevalence of intestinal parasites including microsporidia in human immunodeficiency virus– infected adults in Cameroon: a cross-sectional study. The American Journal of Tropical Medicine and Hygiene. 2006;74(1):162–4.

Kumar SS, Ananthan S, Joyee A. Detection of Enterocytozoon bieneusi (Microsporidia) by polymerase chain reaction (PCR) using species-specific primer in stool samples of HIV patients. Indian J Med Res. 2005;121(4):215–9.

Lono A, Kumar S, Chye TT. Detection of microsporidia in local HIV-positive population in Malaysia. Transactions of the Royal Society of Tropical Medicine and Hygiene. 2011;105(7):409–13.

Tumwine JK, Kekitiinwa A, Bakeera-Kitaka S, Ndeezi G, Downing R, Feng X, et al. Cryptosporidiosis and microsporidiosis in Ugandan children with persistent diarrhea with and without concurrent infection with the human immunodeficiency virus. The American Journal of Tropical Medicine and Hygiene. 2005;73(5):921–5.

Ferreira FM, Bezerra L, Santos MBG, Bernardes RM, Avelino I, Silva MS. Intestinal microsporidiosis: a current infection in HIV-seropositive patients in Portugal. Microbes and Infection. 2001;3(12):1015–9.

Viriyavejakul P, Nintasen R, Punsawad C, Chaisri U, Punpoowong B, Riganti M. High prevalence of Microsporidium infection in HIV-infected patients.Southeast Asian J Trop Med Public Health. 2009 Mar; 40(2):223–8.

Kotler DP, Orenstein JM. Clinical syndromes associated with microsporidiosis. Advances in Parasitology. 1998;40:321–49.

Germani Y, Minssart P, Vohito M, Yassibanda S, Glaziou P, Hocquet D, et al. Etiologies of acute, persistent, and dysenteric diarrheas in adults in Bangui, Central African Republic, in relation to human immunodeficiency virus serostatus. The American Journal of Tropical Medicine and Hygiene. 1998;59(6):1008–14.

Eser B, 체lent. A case of pulmonary Microsporidiasis in an acute myeloblastic leukemia (AML)-M3 patient. Yonsei Medical Journal. 2003;44(1):146–9.

Beatriz L, Isabel L-M, Cristina A, Soledad F, Julio T, del Aguila C. Intestinal microsporidiosis due to Enterocytozoon bieneusi in elderly human immunodeficiency virus– negative patients from Vigo, Spain. Clinical Infectious Diseases. 2002;34(7):918–21.

El Sobky M, El Nahas N. Detection and differentiation between Enterocytozoon bieneusi and Encephalitozoon intestinalis species in cancer patient’s stools using PCR compared with different staining methods. PUJ; 2012.

Lono AR, Kumar S, Chye TT. Incidence of microsporidia in cancer patients. Journal of Gastrointestinal Cancer. 2008;39(1-4):124–9.

Gumbo T, Hobbs RE, Carlyn C, Hall G, Isada CM. Microsporidia infection in transplant patients. Transplantation. 1999;67(3):482–4.

Verweij JJ, ten Hove R, Brienen EA, van Lieshout L. Multiplex detection ofEnterocytozoon bieneusi andEncephalitozoon spp. in fecal samples using real-time PCR. Diagnostic Microbiology and Infectious Disease. 2007;57(2):163–7.

Anane S, Attouchi H. Microsporidiosis: epidemiology, clinical data and therapy. Gastroenterologie Clinique et Biologique. 2010;34(8):450–64.

51.Thellier M, Breton J. Enterocytozoon bieneusi in human and animals, focus on laboratory identification and molecular epidemiology. Parasite. 2008;15(3):349–58.

Patil K, De A, Mathur M. Comparison of weber green and ryan blue modified trichrome staining for the diagnosis of microsporidial spores from stool samples of HIV-positive patients with diarrhoea. Indian Journal of Medical Microbiology. 2008;26(4):407-.

Polley SD, Boadi S, Watson J, Curry A, Chiodini PL. Detection and species identification of microsporidial infections using SYBR Green real-time PCR. Journal of Medical Microbiology. 2011;60(4):459–66.

Angela R, Suresh K. Microsporidia in stool from cancer patients. J Med Sci. 2007;1(2):88–90.

Bednarska M, Bajer A, Siński E, Wolska-Kuśnierz B, Samoliński B, Graczyk T. Occurrence of intestinal microsporidia in immunodeficient patients in Poland. Annals of Agricultural and Environmental Medicine: AAEM. 2014;21(2):244–8.

El-Mahallawy H, Zaki MM, El-Arousy M, Shalabi L, Mansour T. Diagnosis of intestinal microsporidiosis in pediatric oncology patients in Egypt using modified acid fast trichrome staining versus PCR. Acta Parasitologica. 2011;56(4):348–52.

Kašičková D, Sak B, Kváč M, Ditrich O. Sources of potentially infectious human microsporidia: molecular characterisation of microsporidia isolates from exotic birds in the Czech Republic, prevalence study and importance of birds in epidemiology of the human microsporidial infections. Veterinary Parasitology. 2009;165(1):125–30.

Snowden K, Logan K, Phalen D. Isolation and characterization of an avian isolate of Encephalitozoon hellem. Parasitology. 2000;121(1):9–14.

Weber R, Bryan RT, Schwartz DA, Owen RL. Human microsporidial infections. Clinical Microbiology Reviews. 1994;7(4):426.


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