Abstract
Giardia lamblia and Cryptosporidium parvum are recognized as the most common protozoan infections in Saudi Arabia. Microscopic examination of stool samples, either direct or concentrated, for the recovery of G. lamblia cysts and trophozoites and C. parvum oocysts is still the most commonly used for the diagnosis of both parasites. We compared the conventional parasitological techniques of iodine-stained wet mount for G. lamblia and Kinyoun's acid-fast for C. parvum against ImmunoCard STAT® Cryptosporidium/Giardia and real-time polymerase chain reaction (PCR) detecting the 18S rRNA gene of G. lamblia and conventional PCR detecting the same gene of C. parvum at a tertiary hospital in Dhahran, Saudi Arabia. Out of 148 stool samples, 19 and 12 true positives were identified for G. lamblia and C. parvum, respectively, using a composite reference standard. In this case, true positives and negatives were considered as those with at least two positive or negative results out of the three tests. Both ImmunoCard STAT! and PCR methods were more sensitive than the microscopic tests of a single stool specimen of 85.7 % (CI = 62.6–96.2 %) and 85.7 % (CI = 56.2–97.5 %) for G. lamblia and C. parvum, respectively. However, specificity of microscopic tests was higher than other techniques for both parasites. Although PCR seems to be most sensitive for both G. lamblia and C. parvum, its low specificity may render its superiority over other techniques. When a single stool sample is used for detection of G. lamblia and C. parvum, better results can be obtained when coupled with serological testing. Although PCR is the most sensitive method for the detection of both G. lamblia and C. parvum, its use requires attention in relation to the increased possible false positives.
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References
Al-Briken FA, Amin A, Beeching NJ, Hommel M, Hart CA (2003) Detection of Cryptosporidium amongst diarrheic and asymptomatic children in Jeddah, Saudi Arabia. Ann Trop Med Parasitol 97(5):505–510
Alonzo TA, Pepe MS (1999) Using a combination of reference tests to assess the accuracy of a new diagnostic test. Stat Med 18:2987–3003
Ash LR, Orihel TC (1987) Parasites: a guide to laboratory procedures and identification. American Society of Clinical Pathologists Press, Chicago, Ill
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (2005) Current protocols in molecular biology. Wiley, Hoboken
Banoo S, Bell D, Bossuyt P, Herring A, Mabey D, Poole F, Smith PG, Sriram N, Wongsrichanalai C, Linke R, O’Brien R, Perkins M, Cunningham J, Matsoso P, Nathanson CM, Olliaro P, Peeling RW, Ramsay A (2006) Evaluation of diagnostic tests for infectious diseases: general principles. Nat Rev Microbiol 4(12):S20–S32
Birkhead G, Vogt RL (1989) Epidemiologic surveillance for endemic Giardia lamblia infection in Vermont. The roles of waterborne and person-to-person transmission. Am J Epidemiol 129(4):762–768
Bolbol AS (1992) Cryptosporidiosis in young children suffering from diarrhea in Riyadh, Saudi Arabia. J Hyg Epidemiol Microbiol Immunol 36:396–400
Branda JA, Tai-Yuan DL, Rosenberg ES, Halpern EF, Ferraro MJ (2006) A rational approach to the stool ova and parasite examination. Clin Infect Dis 42:972–978
Cartwright CP (1999) Utility of multiple stool specimen ova and parasite examinations in a high prevalence setting. J Clin Microbiol 37:2408–2411
Clinical and Laboratory Standard Institute (CLSI) (2005) Procedures for the recovery and identification of parasites from the intestinal tract; approved guidelines—second edition. CLSI Document M28-A2 (ISBN 1-56238-572-0).
Danciger M, Lopez M (1975) Numbers of Giardia in the feces of infected children. AmJTrop Med Hyg 24(2):237–242
Dib HH, Lu SQ, Wen SF (2008) Prevalence of Giardia lamblia with or without diarrhea in South East, South East Asia and the Far East. Parasitol Res 103(2):239–251
El-Badry AA, Al-Ali KH, Mahrous AS (2010) Molecular identification & prevalence of Giardia lamblia & Cryptosporidium in dudenal aspirate in Al-Madinah. Journal of Medicine and Biomedical Science. ISSN: 2078–0273: 47–52
Elgun G, Koltas IS (2011) Investigation of Cryptosporidium spp. antigen by ELISA method in stool specimens obtained from patients with diarrhea. Parasitol Res 108(2):395–397
El-Moamly AA, El-Sweify MA (2012) ImmunoCard STAT! cartridge antigen detection assay compared to microplate enzyme immunoassay and modified Kinyoun's acid-fast staining technique for detection of Cryptosporidium in fecal specimens. Parasitol Res 110(2):1034–1041
Garcia LS, Shum AC, Bruckner DA (1992) Evaluation of a new monoclonal antibody combination reagent for direct fluorescence detection of Giardia cysts and Cryptosporidium oocysts in human fecal specimens. J Clin Microbiol 30:3255–3257
Garcia LS, Shimizu RY (1997) Evaluation of nine immunoassay kits (enzyme immunoassay and direct fluorescence) for detection of Giardia lamblia and Cryptosporidium parvum in human fecal specimens. J Clin Microbiol 35:1526–1529
Garcia LS, Shimizu RY (2000) Detection of Giardia lamblia and Cryptosporidium parvum antigens in human fecal specimens using the ColorPAC combination rapid solid-phase qualitative immunochromatographic assay. J Clin Microbio 38:1267–1268
Garcia LS, Shimizu RY, Novak S, Carroll M, Chan F (2003) Commercial assay for detection of Giardia lamblia and Cryptosporidium parvum antigens in human fecal specimens by rapid solid-phase qualitative immunochromatography. J Clin Microbiol 41:209–212
Hanson KL, Cartwright CP (2001) Use of an enzyme immunoassay does not eliminate the need to analyze multiple stool specimens for sensitive detection of Giardia lamblia. J Clin Microbiol 39:474–477
Johnston SP, Ballard MM, Beach MJ, Causer L, Wilkens PP (2003) Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens. J Clin Microbiol 41:623–626
Khan ZH, Namnyak SS, Al-Jama AA, Madan I (1988) Prevalence of cryptosporidiosis in Damam and Alkhobar. Saudi Arabia Ann Trop Paediatr 8:170–172
Klein D (2002) Quantification using real-time PCR technology: applications and limitations. Trends Mol Med 8:257–260
Magi B, Canocchi V, Tordini G, Cellesi C, Barberi A (2005) Cryptosporidium infection: diagnostic techniques. Parasitol Res 98(2):150–152
Meinhardt PL, Casemore DP, Miller KB (1996) Epidemiological aspects of human cryptosporidiosis and the role of waterborne transmission. Epidemiol Rev 18:118–136
Meyer EA (1990) Human parasitic diseases: giardiasis. In: Meyer EA (ed) Taxonomy and nomenclature. Elsevier, Amsterdam, pp 51–60
Minak J, Kabir M, Mahmud I, Liu Y, Liu L, Haque R, Petri WA (2012) Evaluation of rapid antigen point-of-care tests for detection of Giardia and Cryptosporidium species in human fecal specimens. J Clin Microbiol 50:154–156
Morgan UM, Pallant L, Dwyer BW, Forbes DA, Rich G, Thompson RCA (1998) Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial. J Clin Microbiol 36:995–998
Ndao M (2010) Diagnosis of parasitic diseases: old and new approaches. Interdisc Perspect on Infect Dis 106(5):1127–1134
Newman RD, Jaeger KL, Wuhib T, Lima AAM, Guerrant RL, Sears CL (1993) Evaluation of an antigen capture enzyme-linked immunosorbent assay for detection of Cryptosporidium oocysts. J Clin Microbiol 31:2080–2084
Omar MS, Abu-Zeid HAH, Mahfouz AAR (1991) Intestinal parasitic infections in schoolchildren of Abha (Asir), Saudi Arabia. Acta Trop 48(3):195–202
Omar MS, Mahfouz AA, Abdel-Moneim M (1995) The relationship of water sources and other determinants to prevalence of intestinal protozoal infections in a rural community of Saudi Arabia. J of Community Health 20(5):433–440
Oster N, Gehrig-Feistel H, Jung H, Kammer J, McLean JE, Lanzer M (2006) Evaluation of the immunochromatographic CORIS Giardia-Strip test for rapid diagnosis of Giardia lamblia. Eur J Clin Microbiol Infect Dis 25(2):112–115
Pincus MR (1996) Interpreting laboratory results: reference values and decision making. In: Hery JB (ed) Clinical diagnosis and management by laboratory methods. Saunders, Philadelphia, pp 76–77
Ridley DS, Hawgood BC (1956) The value of formol-ether concentration of faecal cysts and ova. J Clin Pathol 9(1):74–76
Robinson TJ, Cebelinski EA, Taylor C, Smith KE (2010) Evaluation of the positive predictive value of rapid assays used by clinical laboratories in Minnesota for the diagnosis of cryptosporidiosis. Clin Infect Dis 50(8):e53–e55
Schuurman T, Lankamp P, van Belkum A, Kooistra-Smid M, van Zwet A (2007) Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens. Clin Microbiol Infect 13:1186–1191
Verweij JJ, Pit DSS, Van Lieshout L, Baeta SM, Dery GD, Gasser RB, Polderman AM (2001) Determining the prevalence of Oesophagostomum bifurcum and Necator americanus infections using specific PCR amplification of DNA from faecal samples. Trop Med Int Health 6:726–731
Wybo I, Piérard D, Reynders M, Breynaert J, Covens L, Cnudde D, Lauwers S (2005) Evaluation of ImmunoCard STAT! immunoassay in the detection of Giardia lamblia and Cryptosporidium parvum specific antigens. 15th European Congress of Clinical Microbiology and Infectious Diseases Copenhagen/Denmark, April 2–5, 2005. Abstract number: 1134_03_261
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This work was supported by the Prince Sultan Military College of Health Sciences. We thank the College Director Maj. Gen. Saleh H Al-Shayea. Col/Yasser Al Neam and Mr. Abdulla Al Shehri helped us a lot in procurement of supplies and samples, to whom we are indebted.
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Elsafi, S.H., Al-Maqati, T.N., Hussein, M.I. et al. Comparison of microscopy, rapid immunoassay, and molecular techniques for the detection of Giardia lamblia and Cryptosporidium parvum . Parasitol Res 112, 1641–1646 (2013). https://doi.org/10.1007/s00436-013-3319-1
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DOI: https://doi.org/10.1007/s00436-013-3319-1