Comparison of the Triage Micro Parasite Panel and Microscopy for the Detection of Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Cryptosporidium parvum in Stool Samples Collected in Kenya
Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are three of the most important parasitic causes of acute diarrhea worldwide. Laboratory diagnosis of these parasites is usually done by ova and parasite examination (O&P examination) via microscopy. The sensitivity and specificity of O&P examination varies among laboratories and can be labor intensive and time consuming. The Triage Micro Parasite Panel (BioSite, San Diego, California) is an enzyme immunoassay kit that can detect E. histolytica/E. dispar, G. lamblia, and C. parvum simultaneously using fresh or frozen stool. The present study evaluated the Triage Micro Parasite Panel in detecting E. histolytica/E. dispar, G. lamblia, and C. parvum compared to O&P examination in 266 stool samples collected at medical facilities in Kenya. The sensitivity and specificity results for the Triage Micro Parasite Panel were: for E. histolytica/E. dispar: 100%, 100%, G. lamblia: 100%, 100% and C. parvum: 73%, 100%. There was no evidence of cross reactivity using the kit with other parasites identified in the stool specimens. These results indicate that the Triage Micro Parasite Panel is a highly sensitive kit that can be used for screening purposes in large scale studies or outbreak investigations or as a possible alternative to O&P examination. 1. Introduction Diarrheal disease is a major cause of morbidity and mortality worldwide particularly in developing countries where poor sanitary and hygienic conditions exist [1–4]. The most common parasitic causes of acute diarrhea are the intestinal protozoa of which Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are considered the most important [5, 6]. Detection of trophozoites, cysts, or oocysts in fresh or preserved stool specimens using microscopy (ova and parasite examination (O&P)) is the most common method of diagnosis particularly in resource limited countries. Though microscopy is fairly inexpensive, it can be time consuming and labor intensive, and diagnosis usually depends on the microscopist’s level of expertise and training [7–10]. Due to the technical variability among laboratory technologists, misidentification of these parasites, particularly E. histolytica, has led to unnecessary or delayed treatment [11, 12]. In addition, previous studies have shown that excretion of trophozoites, cysts, or oocysts in the feces can be intermittent and sporadic from day to day and therefore could lead to missed infections due to the low numbers of the diagnostic stages in the stool sample [13, 14]. For these reasons, several rapid
References
[1]
C. J. L. Murray and A. D. Lopez, “Mortality by cause for eight regions of the world: global burden of disease study,” The Lancet, vol. 349, no. 9061, pp. 1269–1276, 1997.
[2]
R. L. Guerrant, J. M. Hughes, N. L. Lima, and J. Crane, “Diarrhea in developed and developing countries: magnitude, special settings, and etiologies,” Reviews of Infectious Diseases, vol. 12, supplement 1, pp. S41–S50, 1990.
[3]
M. Kosek, C. Bern, and R. L. Guerrant, “The global burden of diarrhoeal disease, as estimated from studies published between 1992 and 2000,” Bulletin of the World Health Organization, vol. 81, no. 3, pp. 197–204, 2003.
[4]
C. L. F. Walker, M. J. Aryee, C. Boschi-Pinto, R. E. Black, et al., “Estimating diarrhea mortality among young children in low and middle income countries,” PLoS One, vol. 7, no. 1, Article ID e29151, 2012.
[5]
C. Kourenti, P. Karanis, and H. Smith, “Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt,” Journal of Water and Health, vol. 5, no. 1, pp. 1–38, 2007.
[6]
P. C. Okhuysen, “Traveler's diarrhea due to intestinal protozoa,” Clinical Infectious Diseases, vol. 33, no. 1, pp. 110–114, 2001.
[7]
T. G. Mank, J. O. M. Zaat, A. M. Deelder, J. T. M. Van Eijk, and A. M. Polderman, “Sensitivity of microscopy versus enzyme immunoassay in the laboratory diagnosis of giardiasis,” European Journal of Clinical Microbiology and Infectious Diseases, vol. 16, no. 8, pp. 615–619, 1997.
[8]
M. M. Marshall, D. Naumovitz, Y. Ortega, and C. R. Sterling, “Waterborne protozoan pathogens,” Clinical Microbiology Reviews, vol. 10, no. 1, pp. 67–85, 1997.
[9]
S. P. Johnston, M. M. Ballard, M. J. Beach, L. Causer, and P. P. Wilkins, “Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens,” Journal of Clinical Microbiology, vol. 41, no. 2, pp. 623–626, 2003.
[10]
R. A. Nesbitt, F. W. Mosha, H. A. Katki, M. Ashraf, C. Assenga, and C. M. Lee, “Amebiasis and comparison of microscopy to ELISA technique in detection of Entamoeba histolytica and Entamoeba dispar,” Journal of the National Medical Association, vol. 96, no. 5, pp. 671–677, 2004.
[11]
A. Kebede, J. Verweij, W. Dorigo-Zetsma et al., “Overdiagnosis of amoebiasis in the absence of Entamoeba histolytica among patients presenting with diarrhoea in Wonji and Akaki, Ethiopia,” Transactions of the Royal Society of Tropical Medicine and Hygiene, vol. 97, no. 3, pp. 305–307, 2003.
[12]
A. Kebede, J. J. Verweij, B. Petros, and A. M. Polderman, “Short communication: misleading microscopy in amoebiasis,” Tropical Medicine and International Health, vol. 9, no. 5, pp. 651–652, 2004.
[13]
M. Danciger and M. Lopez, “Numbers of Giardia in the feces of infected children,” American Journal of Tropical Medicine and Hygiene, vol. 24, no. 2, pp. 237–242, 1975.
[14]
R. D. Newman, K. L. Jaeger, T. Wuhib, A. A. M. Lima, R. L. Guerrant, and C. L. Sears, “Evaluation of an antigen capture enzyme-linked immunosorbent assay for detection of Cryptosporidium oocysts,” Journal of Clinical Microbiology, vol. 31, no. 8, pp. 2080–2084, 1993.
[15]
L. S. Garcia and R. Y. Shimizu, “Evaluation of nine immunoassay kits (enzyme immunoassay and direct fluorescence) for detection of Giardia lamblia and Cryptosporidium parvum in human fecal specimens,” Journal of Clinical Microbiology, vol. 35, no. 6, pp. 1526–1529, 1997.
[16]
R. Haque, I. K. M. Ali, S. Akther, and W. A. Petri, “Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection,” Journal of Clinical Microbiology, vol. 36, no. 2, pp. 449–452, 1998.
[17]
S. Buss, M. Kabir, W. A. Petri, and R. Haque, “Comparison of two immunoassays for detection of Entamoeba histolytica,” Journal of Clinical Microbiology, vol. 46, no. 8, pp. 2778–2779, 2008.
[18]
L. S. Garcia, R. Y. Shimizu, and C. N. Bernard, “Detection of Giardia lamblia, Entamoeba histolytica/Entamoeba dispar, and Cryptosporidium parvum antigens in human fecal specimens using the triage parasite panel enzyme immunoassay,” Journal of Clinical Microbiology, vol. 38, no. 9, pp. 3337–3340, 2000.
[19]
S. E. Sharp, C. A. Suarez, Y. Duran, and R. J. Poppiti, “Evaluation of the triage micro parasite panel for detection of Giardia lamblia, Entamoeba histolytica/Entamoeba dispar, and Cryptosporidium parvum in patient stool specimens,” Journal of Clinical Microbiology, vol. 39, no. 1, pp. 332–334, 2001.
[20]
B. Leiva, M. Lebbad, J. Winiecka-Krusnell, I. Altamirano, A. Tellez, and E. Linder, “Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study,” Archives of Medical Research, vol. 37, no. 4, pp. 529–534, 2006.
[21]
M. R. Gaafar, “Evaluation of enzyme immunoassay techniques for diagnosis of the most common intestinal protozoa in fecal samples,” International Journal of Infectious Diseases, vol. 15, no. 8, pp. e541–e544, 2011.
[22]
L. S. Garcia, Diagnostic Medical Parasitology, American Society for Microbiology, Washington, DC, USA, 4th edition, 2001.
[23]
R. G. Knapp and M. C. Miller, Clinical Epidemiology and Biostatistics, Harwal, Baltimore, Md, USA, 1992.
[24]
J. A. Platts-Mills, J. Darwin, E. R. Houpt, et al., “Molecular diagnosis of diarrhea: current status and future potential,” Current Infectious Disease Reports, vol. 14, no. 1, pp. 41–46, 2012.
[25]
A. M. van Eijk, J. T. Brooks, P. M. Adcock et al., “Diarrhea in children less than two years of age with known HIV status in Kisumu, Kenya,” International Journal of Infectious Diseases, vol. 14, no. 3, pp. e220–e225, 2010.
[26]
R. N. Chunge, N. Nagelkerke, P. N. Karumba et al., “Longitudinal study of young children in Kenya: intestinal parasitic infection with special reference to Giarida lamblia, its prevalence, incidence and duration, and its association with diarrhoea and with other parasites,” Acta Tropica, vol. 50, no. 1, pp. 39–49, 1991.
[27]
W. Gatei, C. N. Wamae, C. Mbae et al., “Cryptosporidiosis: prevalence, genotype analysis, and symptoms associated with infections in children in Kenya,” American Journal of Tropical Medicine and Hygiene, vol. 75, no. 1, pp. 78–82, 2006.
[28]
D. R. Pillai, J. S. Keystone, D. C. Sheppard, J. D. MacLean, D. W. MacPherson, and K. C. Kain, “Entamoeba histolytica and Entamoeba dispar: epidemiology and comparison of diagnostic methods in a setting of nonendemicity,” Clinical Infectious Diseases, vol. 29, no. 5, pp. 1315–1318, 1999.
[29]
R. M. Chalmers, “Cryptosporidium: from laboratory diagnosis to surveillance and outbreaks,” Parasite, vol. 15, no. 3, pp. 372–378, 2008.
[30]
H. L. DuPont, C. L. Chappell, C. R. Sterling, P. C. Okhuysen, J. B. Rose, and W. Jakubowski, “The infectivity of Cryptosporidium parvum in healthy volunteers,” New England Journal of Medicine, vol. 332, no. 13, pp. 855–859, 1995.
[31]
L. S. Diamond and C. G. Clark, “A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925,” The Journal of Eukaryotic Microbiology, vol. 40, no. 3, pp. 340–344, 1993.
[32]
P. Gonin and L. Trudel, “Detection and differentiation of Entamoeba histolytica and Entamoeba dispar isolates in clinical samples by PCR and enzyme-linked immunosorbent assay,” Journal of Clinical Microbiology, vol. 41, no. 1, pp. 237–241, 2003.
[33]
P. G. Sargeaunt and J. E. Williams, “Electrophoretic isoenzyme patterns of the pathogenic and non-pathogenic intestinal amoebae of man,” Transactions of the Royal Society of Tropical Medicine and Hygiene, vol. 73, no. 2, pp. 225–227, 1979.
[34]
M. Tanyuksel and W. A. Petri Jr, “Laboratory diagnosis of amebiasis,” Clinical Microbiology Reviews, vol. 16, no. 4, pp. 713–729, 2003.
[35]
N. C. Christy, J. D. Hencke, A. Escueta-De Cadiz, et al., “Multisite performance evaluation of an enzyme-linked immunosorbent assay for detection of giardia, Cryptosporidium, and Entamoeba histolytica antigens in human stool,” Journal of Clinical Microbiology, vol. 50, no. 5, pp. 1762–1763, 2012.
[36]
R. Haque, S. Roy, A. Siddique et al., “Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp,” American Journal of Tropical Medicine and Hygiene, vol. 76, no. 4, pp. 713–717, 2007.