CoproELISA Entamoeba Assay Kit

$390.00

The Eagle Bioscience’s CoproELISA Entamoeba Assay Kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for detection of Entamoeba antigens in unpreserved human fecal specimens collected from patients with gastrointestinal symptoms. The test can be used for fecal specimens submitted for routine clinical testing from adults or children. The Eagle Biosciences CoproELISA Entamoeba Assay Kit is for Research Use Only and is not intended for diagnostic or therapeutic purposes.

CoproELISA Entamoeba Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: Cut-Off Control
Dynamic Range: 0-180 pmol/l
Incubation Time: 3 hours
Sample Type: Stool
Sample Size: 0.1 to 0.3g


Additional Information

Assay Background

Entamoeba histolytica/dispar (E. histolytica/dispar) are intestinal protozoan parasites that are transferred via the faecal–oral route and infect up to 10% of the human population (1). Entamoeba infections are most common in the developing world and are associated with poor sanitation where the Entamoeba cysts are transmitted from contaminated food and water. After ingestion trophozoites the Entamoeba vegetative form are released from their cyst capsules into the small intestine. Trophozoites which survive strictly in the host intestine and fresh stool migrate to the large intestine where they live and multiply by binary fission. Trophozoites of E. dispar are noninvasive and reside in the intestine while those of E. histolytica can invade the intestinal mucosa causing human amoebiasis (1 and 2). Symptoms of amoebiasis can range from mild diarrhea to dysentery with blood and mucus in the stool. While most E. histolytica infected people remain asymptomatic, without proper treatment, this disease can lead to severe complications, including abscess of vital organs, most commonly in the liver, and in some cases in the lungs, and the brain (3). It is estimated that E. histolytica, represents 10% of human Entamoeba infections which cause the death of between 40,000 to 100,000 people every year (4). It is, therefore, important to diagnose the infection and provide appropriate treatment. The most common treatment for amoebiasis is the administration of antibiotics, such as metronidazole and tinidazole.

Assay Principle

Plates are coated with specific polyclonal antibodies directed against Entamoeba antigens. Fecal sample to be tested is diluted in stool diluent and incubated with the pre-coated plate. In this step Entamoeba antigens are bound to the immobilized antibodies. Non-specific antigens are removed by washing. Anti- Entamoeba monoclonal antibody conjugated to horseradish peroxidase (HRP) is added and incubated. In this step the HRP-conjugate is bound to the pre-bound antigen-antibody complex. Unbound conjugate is removed by washing. Upon the addition of TMB-substrate, the substrate is hydrolyzed by the peroxidase, yielding a blue solution of the reduced substrate. Upon the addition of the stop solution, the blue color turns yellow and should be read by an ELISA reader at a wavelength of 450/620 nm. The absorbance is proportional to the number of Entamoeba cells present in the sample.

Cross Reaction

The CoproELISA Entamoeba test was evaluated using stool specimens defined as positive for various gastrointestinal pathogens. No cross-reactivity of interference by mixed infection with any of the pathogens listed: Blastocystis,  C. difficile, Cryptosporidium, E. coli, E. nana, D. fragilis, G. lamblia, Salmonella spp.

References

  1. WHO (1997) News and activities. Entamoeba taxonomy. Bull World Health Organ 75: 291–293.
  2. Diamond LS and Clar. A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925. k CG. A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925.  J Eukaryot Microbiol. 40(3): 340-4 (1993).
  3. Tanyuksel M and Petri WA., Laboratory Diagnosis of Amebiasis. Jr. Clin Microbiol Rev. 2003 October; 16(4): 713-729.
  4. Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J. Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev. 2007 Jul;20(3):511-32.

Manual

Product Manual