Abstract
Diarrhea is a debilitating condition in HIV infected individuals and with the finding that almost 1/4 cases of diarrhea in HIV are due to microsporidia, there is a dire need to institute measures for its detection on a regular basis. Keeping this in mind the study aims to determine the burden of intestinal microsporidiosis in HIV seropositive patients presenting with and without diarrhea and to compare the ability of microscopy and PCR in its detection.
The study group consisted of 120 patients divided into four groups HIV seropositive with/without diarrhea, and HIV seronegative with/without diarrhea. Performance of four staining techniques including Modified Trichrome, Calcofluor White, Gram Chromotrope and Quick hot Gram Chromotrope stains were evaluated against PCR in diagnosing enteric microsporidiosis from stool samples.
Overall prevalence of intestinal microsporidiosis was 10.83%. The same for HIV seropositive patients with diarrhea was 23.33%, HIV seropositive patients without diarrhea and in immune-competent hosts with diarrhea was 10% each. Enterocytozoon bieneusi was found to predominate. Calcofluor white stain detected maximum microsporidia in stool samples (76.92%), followed by Modified Trichrome stain (61.5%), PCR (46.15%) and Gram Chromotrope and Quick hot Gram Chromotrope stains (38.4% each). PCR exhibited the best performance with a sensitivity and specificity of 100%. Our data suggests screening of stool samples with either Modified Trichrome or Calcofluor white stain followed by PCR confirmation thus leading to maximum detection along with speciation for complete cure.
References
Agholi M, Hatam GR, Motazedian MH (2013) HIV/AIDS-associated opportunistic protozoal diarrhea. AIDS Res Hum Retrovir 29(1):35–41
Akinbo FO, Okaka CE, Omoregie R (2010) Prevalence of intestinal parasitic infections among HIV patients in Benin City, Nigeria. Libyan J Med 5:5506
Anane S, Attouchi H (2010) Microsporidiosis: epidemiology, clinical data and therapy. Gastroenterol Clin Biol 34:450–464
Barratt JL, Harkness J, Marriott D, Ellis JT, Stark D (2010) Importance of nonenteric protozoan infections in immunocompromised people. Clin Microbiol Rev 23(4):795–836
Bryan RT, Schwartz DA (1999) Epidemiology of microsporidiosis. In: Witner M, Weiss LM (eds) The microsporidia and microsporidiosis. ASM Press, Washington, DC, pp 502–516
Candramathi S, Suresh K, Anita ZA, Kuppusamy UR (2012) Infections of blastocystis hominis and microsporidia in cancer patients: are they opportunistic? Trans R Soc Trop Med Hyg 106:267–269
Cimerman S, Cimerman B, Lewi DS (1999) Prevalence of intestinal parasitic infections in patients with acquired immunodeficiency syndrome in Brazil. Int J Infect Dis 3(4):203–206
Coyle CM, Wittner M, Kotler DP, Noyer C, Orenstein JM, Tanowitz HB, Weiss LM (1996) Prevalence of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon (Septata) intestinalis among patients with AIDS-related diarrhea: determination by polymerase chain reaction to the microsporidial small-subunit rRNA gene. Clin Infect Dis 23(5):1002–1006
Dalvi S, Mehta P, Koticha A, Gita N (2006) Microsporidia as an emerging cause of parasitic diarrhea in HIV seropositive individuals in Mumbai. Bombay Hosp J 48:592–597
Didier ES (2005) Microsporidiosis: an emerging and opportunistic infection in humans and animals. Acta Trop 94:61–76
Didier ES, Weiss LM (2006) Microsporidiosis: current status. Curr Opin Infect Dis 19(5):485–492
Espern A, Morio F, Miegeville M, Illa H, Abdoulaye M, Meyssonnier V, Adehossi E, Lejeune A, Cam PD, Besse B, Gay-Andrieu F (2007) 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. J Clin Microbiol 45(9):2999–3002
Fedorko DP, Hijazi YM (1996) Application of molecular techniques to the diagnosis of microsporidial infection. Emerg Infect Dis 2:183–191
Fournier S, Liguory O, Sarfati C, David-Ouaknine F, Derouin F, Decazes J, Molina J (2000) Disseminated infection due to Encephalitozoon cuniculi in a patient with AIDS: case report and review. HIV Med 1(3):155–161
Garcia LS (2002) Laboratory identification of the microsporidia. J Clin Microbiol 40:1892–1901
Ghoshal U, Khanduja S, Pant P, Prasad KN, Dhole TN, Sharma RK, Ghoshal UC (2015a) Intestinal microsporidiosis in renal transplant recipients: prevalence, predictors of occurrence and genetic characterization. Indian J Med Microbiol 33:357–363
Ghoshal U, Khanduja S, Agarwal V, Dhole TN, Ghoshal UC (2015b) Comparative evaluation of staining techniques and polymerase chain reaction for diagnosis of intestinal microsporidiosis in immunocompromised patients. Trop Parasitol 5:101–105
Ghoshal U, Dey A, Ranjan P, Khanduja S, Agarwal V, Ghoshal UC (2016) Identification of opportunistic enteric parasites among immunocompetent patients with diarrhea from Northern India and genetic characterization of Cryptosporidium and Microsporidia. Indian J Med Microbiol 34:60–66
Gupta M, Sinha M, Raizada N (2008) Opportunistic intestinal protozoan parasitic infection in HIV positive patients in Jamnagar, Gujrat. SAARC J Tuber Lung Dis HIV/AIDS 5(1):21–24
Khan IA, Moretto M, Weiss LM (2001) Immune response to Encephalitozoon cuniculi infection. Microbes Infect 3:401–405
Kulkarni S, Pastute S, Chandane M, Risbud A (2011) Performance of microscopy for detection of microsporidial spores from stool samples of HIV infected individuals with diarrhea. Indian J Med Res 134:982–984
Lee SC, Corradi N, Doan S, Dietrich FS, Keeling PJ, Heitman J (2010) Evolution of the sex-related locus and genomic features shared in microsporidia and fungi. PLoS One 5:e10539
Mathis A, Weber R, Deplazes P (2005) Zoonotic potential of the microsporidia. Clin Microbiol Rev 18(3):423–445
Matsubayashi H, Koike T, Mikata T, Hagiwara S (1959) A case of Encephalitozoon like body infection in man. Arch Pathol 67:181–187
Nkinin SW, Asonganyi T, Didier ES, Kaneshiro ES (2007) Microsporidial infection is prevalent in healthy people in Cameroon. J Clin Microbiol 45(9):2841–2846
Raynaud L, Delbac F, Broussolle V, Rabodonirina M, Girault V, Wallon M, Cozon G, Vivares CP, Peyron F (1998) Identification of Encephalitozoon intestinalis in travelers with chronic diarrhea by specific PCR amplification. J Clin Microbiol 36(1):37–40
Saigal K, Sharma A, Sehgal R, Sharma P, Malla N, Khurana S (2013a) Intestinal microsporidiosis in India: a two year study. Parasitol Int 62(1):53–56
Saigal K, Khurana S, Sharma A, Sehgal R, Malla N (2013b) Comparison of staining techniques and multiplex nested PCR for diagnosis of intestinal microsporidiosis. Diag Microbiol Infect Dis 77:248–249
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. J Clin Microbiol 49(3):1064–1070
Samie A, Obi CL, Tzipori S, Weiss LM, Guerrant RL (2007) Microsporidiosis in South Africa: PCR detection in stool samples of HIV positive and HIV negative individuals and school children in Vhembe district, Limpopo Provinces. Trans R Soc Trop Med Hyg 101(6):547–554
Tuli L, Gulati AK, Sundar S, Mohapatra TM (2008) Correlation between CD4 counts of HIV patients and enteric protozoan in different seasons – an experience of a tertiary care hospital in Varanasi (India). BMC Gastroenterol 8:36
Türk S, Dogruman AF, Karaman U, Kustimur S (2012) Investigation of Microsporidia prevalence by different staining methods in cases of diarrhea. Microbiol Bull 46:85–92
Viera AJ, Garrett JM (2005) Understanding inter observer agreement: the kappa statistic. Fam Med 37(5):360–363
Visvesvara GS, Leitch GJ, Pieniazek NJ, Da Silva AJ, Wallace S, Slemenda SB, Weber R, Schwartz DA, Gorelkin L, Wilcox CM (1995) Short term in vitro culture and molecular analysis of the microsporidian, Enterocytozoon bieneusi. J Euk Microbiol 42:506–510
Weber R, Bryan RT, Schwartz DA, Owen RL (1994) Human microsporidial infections. Clin Microbiol Rev 7:426–461
Yakoob J, Abbas Z, Beg MA, Jafri W, Naz S, Khalid A, Khan R (2012) Microsporidial infections due to Encephalitozoon intestinalisin in non-HIV-infected patients with chronic diarrhea. Epidemiol Infect 140:1773–1779
Zhu X, Wittner M, Tanowitz HB, Kotler D, Cali A, Weiss LM (1993) Small subunit rRNA sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction. J Infect Dis 168:1570–1575
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer International Publishing AG
About this chapter
Cite this chapter
Kaushik, S., Saha, R., Das, S., Ramachandran, V., Goel, A. (2017). Pragmatic Combination of Available Diagnostic Tools for Optimal Detection of Intestinal Microsporidia. In: Donelli, G. (eds) Advances in Microbiology, Infectious Diseases and Public Health. Advances in Experimental Medicine and Biology(), vol 1057. Springer, Cham. https://doi.org/10.1007/5584_2017_97
Download citation
DOI: https://doi.org/10.1007/5584_2017_97
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-79016-9
Online ISBN: 978-3-319-79017-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)