Quantitation of Giardia Cysts and Cryptosporidium Oocysts in Fecal Samples by Direct Immunofluorescence Assay

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 1993 Nov 1

PMID 8263179

Citations 19

Authors

L Xiao

R P Herd

Affiliations

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Abstract

The lack of quick, simple, and sensitive quantitative tests has impeded studies on infection patterns and treatment of Giardia spp. and Cryptosporidium spp. A quantitative direct immunofluorescence assay (FA) using a commercial FA kit was developed and evaluated. Recovery rates of the FA for Cryptosporidium oocysts in calf feces seeded with 1,000, 10,000, 100,000, and 1,000,000 oocysts per g were 14.8, 40.8, 84.2, and 78.2%, respectively. Interassay coefficients of variation were 10.6 to 47.1%. Recovery rates of the FA for Giardia cysts in feces seeded with 1,000, 10,000, and 100,000 cysts per g were 76.4, 96.9, and 89.6%, respectively. Interassay coefficients of variation were 7.4 to 22.1%. By comparison, recovery rates of Giardia cyst by sucrose gradient flotation were only 20.5, 51.2, and 42.9%, respectively. Counts of cysts-per-gram obtained by sucrose gradient flotation with samples from calves, lambs, and ewes were only 49.1 to 54.8% of those obtained by the FA. Zinc sulfate flotation detected only 36.4% of infections when there were < or = 1,000 cysts per g. The quantitative FA offers a useful technique for epidemiological and control studies of these two parasites.

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References

Stein E, BOCH J, Heine J, Henkel G . [Course of natural Cryptosporidium infections on 4 cattle-breeding farms]. Berl Munch Tierarztl Wochenschr. 1983; 96(7):222-5. View

Roberts-Thomson I, Stevens D, Mahmoud A, Warren K . Giardiasis in the mouse: an animal model. Gastroenterology. 1976; 71(1):57-61. View

Gasser R, Eckert J, Rohrer L . Isolation of Giardia from Swiss cattle and cultivation of trophozoites in vitro. Parasitol Res. 1987; 73(2):182-3. DOI: 10.1007/BF00536478. View

Gasser R, Eckert J, Rohrer L . Infectivity of Swiss Giardia isolates to jirds and mice, and in vitro cultivation of trophozoites originating from sheep. Parasitol Res. 1987; 74(2):103-11. DOI: 10.1007/BF00536020. View

Iseki M, Maekawa T, Moriya K, Uni S, Takada S . Infectivity of Cryptosporidium muris (strain RN 66) in various laboratory animals. Parasitol Res. 1989; 75(3):218-22. DOI: 10.1007/BF00931279. View

Rusnak J, Hadfield T, Rhodes M, Gaines J . Detection of Cryptosporidium oocysts in human fecal specimens by an indirect immunofluorescence assay with monoclonal antibodies. J Clin Microbiol. 1989; 27(5):1135-6. PMC: 267504. DOI: 10.1128/jcm.27.5.1135-1136.1989. View

Arrowood M, Sterling C . Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection. J Clin Microbiol. 1989; 27(7):1490-5. PMC: 267601. DOI: 10.1128/jcm.27.7.1490-1495.1989. View

Taminelli V, Eckert J, Sydler T, Gottstein B, Corboz L, Hofmann M . [Experimental infection of calves and sheep with bovine Giardia isolates]. Schweiz Arch Tierheilkd. 1989; 131(9):551-64. View

Erlandsen S, Sherlock L, Bemrick W, Ghobrial H, Jakubowski W . Prevalence of Giardia spp. in beaver and muskrat populations in northeastern states and Minnesota: detection of intestinal trophozoites at necropsy provides greater sensitivity than detection of cysts in fecal samples. Appl Environ Microbiol. 1990; 56(1):31-6. PMC: 183246. DOI: 10.1128/aem.56.1.31-36.1990. View

10.

Buret A, Denhollander N, Wallis P, Befus D, Olson M . Zoonotic potential of giardiasis in domestic ruminants. J Infect Dis. 1990; 162(1):231-7. DOI: 10.1093/infdis/162.1.231. View

11.

Asahi H, Koyama T, Arai H, Funakoshi Y, Yamaura H, SHIRASAKA R . Biological nature of Cryptosporidium sp. isolated from a cat. Parasitol Res. 1991; 77(3):237-40. DOI: 10.1007/BF00930864. View

12.

Weber R, Bryan R, Bishop H, Wahlquist S, Sullivan J, Juranek D . Threshold of detection of Cryptosporidium oocysts in human stool specimens: evidence for low sensitivity of current diagnostic methods. J Clin Microbiol. 1991; 29(7):1323-7. PMC: 270109. DOI: 10.1128/jcm.29.7.1323-1327.1991. View

13.

Cozon G, Cannella D, Biron F, Piens M, Jeannin M, Revillard J . Cryptosporidium parvum sporozoite staining by propidium iodide. Int J Parasitol. 1992; 22(3):385-9. DOI: 10.1016/s0020-7519(05)80018-9. View

14.

Weber R, Bryan R, Juranek D . Improved stool concentration procedure for detection of Cryptosporidium oocysts in fecal specimens. J Clin Microbiol. 1992; 30(11):2869-73. PMC: 270544. DOI: 10.1128/jcm.30.11.2869-2873.1992. View

15.

Garcia L, Shum A, Bruckner D . 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. 1992; 30(12):3255-7. PMC: 270643. DOI: 10.1128/jcm.30.12.3255-3257.1992. View

16.

Mtambo M, Nash A, Blewett D, Wright S . Comparison of staining and concentration techniques for detection of Cryptosporidium oocysts in cat faecal specimens. Vet Parasitol. 1992; 45(1-2):49-57. DOI: 10.1016/0304-4017(92)90026-6. View

17.

Goodgame R, Genta R, White A, Chappell C . Intensity of infection in AIDS-associated cryptosporidiosis. J Infect Dis. 1993; 167(3):704-9. DOI: 10.1093/infdis/167.3.704. View

18.

Garcia L, Brewer T, Bruckner D . Fluorescence detection of Cryptosporidium oocysts in human fecal specimens by using monoclonal antibodies. J Clin Microbiol. 1987; 25(1):119-21. PMC: 265837. DOI: 10.1128/jcm.25.1.119-121.1987. View