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Human Anti-Giardia Iamblia IgA ELISA Assay Kit

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$620.00
SKU:
GIA31-K01

 Product Description

Package insert view pdf

 

 

Human Anti-Giardia Iamblia IgA ELISA Assay Kit: 
For Research Use Only
Size:  1x96 wells
Sensitivity:  1 U/mL
Dynamic Range: 0 - 100 U/mL
Incubation Time:  2 hours 
Sample Type:  Serum, plasma

Sample Size: 10 µL
Controls Included


Intended Use

This Eagle Biosciences Fecal Giardia Iamblia Antigen ELISA (enzyme linked immunosorbent assay) kit is intended for the qualitative detection of Giardia lamblia antigen in feces. The assay is a useful tool in the diagnosis of active Giardia lamblia infection in acute or chronic gastroenteritis.  The Eagle Biosciences Fecal Giardia Iamblia ELISA Assay kit is intended for research use only and not intended for diagnostic procedures.

Assay Principle

This Eagle Biosciences Human Anti-Giardia Iamblia IgA ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human anti-Giardia lamblia IgA in test specimen. The assay utilizes the microplate-based enzyme immunoassay technique by coating highly purified and inactive Giardia lamblia antigen onto the wall of microtiter well.

Assay standards, controls and unknown specimen are added to microtiter wells of microplate that was coated with a highly-purified Giardia lamblia antigen on its wall. The Giardia lamblia antigen will be bound to the antibody in the liquid standards, controls and test samples. The unbound matrices are washed away and a HRP-conjugated antibody which specifically recognizes the specific subtype of human antibody (IgA) is added for further immunoreactions.  After an incubation period, an immunocomplex of “Giardia lamblia Antigen – human Anti-Giardia IgA– HRP-conjugated Anti-hIgA Antibody” is formed if the human anti-Giardia IgA is present in the test sample. The unbound tracer antibody and other protein or buffer matrix are removed in the subsequent washing step. HRP-conjugated tracer antibody bound to the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the tracer antibody bound to the wall of each microtiter well is directly proportional to the amount of human Anti-Giardia lamblia IgA level in each test specimen.

  1. Place a sufficient number of Giardia antigen-coated microwell strips in a frame.   
  2. Add 100 µL of standards, controls and diluted patient serum samples into the designated microwell.
  3. Cover the plate with one plate sealer.
  4. Incubate plate at room temperature for 1 hour.
  5. Prepare working anti-hIgA Tracer Antibody Working Solution by 1:21 fold dilution of the tracer antibody with the Tracer Antibody Diluent. For each strip, it is required to mix 1 mL of Tracer Antibody Diluent with 50 µL of IgA Tracer Antibody in a clean test tube.
  6. Remove the plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL to 400 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  7. Add 100 µL of above diluted tracer antibody working solution to each of the wells.
  8. Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
  9. Incubate plate at room temperature for 30 minutes.
  10. Remove the plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL to 400 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  11. Add 100 µL of ELISA HRP Substrate into each of the wells.
  12. Cover the plate with a new plate sealer and also with aluminum foil to avoid exposure to light.
  13. Incubate plate at room temperature for 10 minutes.
  14. Remove the aluminum foil and plate sealer.  Add 100 µL of ELISA Stop Solution  into each of the wells. Mix gently.
  15. Read the absorbance at 450 nm within 10 minutes in a microplate reader.

Assay Background

Giardia lamblia (also known as Giardia intestinalis) has a characteristic tear-drop shape and measures 10-15 µm in length. It has twin nuclei and an adhesive disk which is a rigid structure reinforced by supelicular microtubules. There are two median bodies of unknown function, but their shape is important for differentiating between species. There are 4 pairs of flagella, one anterior pair, two posterior pairs and a caudal pair. These organisms have no mitochondria, endoplasmic reticulum, golgi, or lysosomes. Giardia has a two-stage life cycle consisting of trophozoite and cyst. The life cycle begins with ingested cysts, which release trophozoites (10-20 µm x 5-15 µm) in the duodenum. These trophozoites attach to the surface of the intestinal epithelium using a ventral sucking disk and then reproduce by binary fission. The trigger for encystment is unclear, but the process results in the inactive, environmentally resistant form of Giardia -- a cyst (11-14 µm x 7-10 µm) that is excreted in feces. Giardiasis is a diarrheal illness caused by Giardia lamblia, after ingestion of Giardia cysts. Once a person has been infected with Giardia, the parasite lives in the intestine and is passed in the stool. Millions of germs can be released in a bowel movement from an infected human or animal. Giardia is found in soil, food, water, or surfaces that have been contaminated with the feces from infected humans or animals. Because the parasite is protected by an outer shell, it can survive outside the body and in the environment for long periods of time. Because it is spread world-wide, Giardia lamblia has become one of the most important causes of chronic diarrheas. About 15-20% of children under age ten years and 19% of male homosexuals have been infected. Giardia infection can cause a variety of intestinal symptoms either acute or chronic, which include diarrhea, gas or flatulence, greasy stools that tend to float, stomach cramps, upset stomach or nausea. These symptoms may lead to weight loss and dehydration. Some people with giardiasis have no symptoms at all. Those asymptomatic cases still shed Giardia cysts. Generally, symptoms of giardiasis begin 1 to 2 weeks after becoming infected and they may last 2 to 6 weeks.  Despite the fact that Giardia is essentially a luminal pathogen in the gut it does evoke both systemic and local immune responses. Current between serum and secretory antibody responses remains unclear; the presence of anti-Giardia antibodies in serum would be in any way indicative of the development of protective immunity. Evidence emphasizes the importance of secretory antibody for clearance of the pathogen, although other cell-mediated effector mechanisms are also likely to be involved.  Recent studies have found that about 86% of infected patients develops serum antibody (IgG and IgA) against Giardia lamblia. Determination of human anti-giardia antibody may contribute to the aid of clinical diagnosis and understand the status of immune response for each individual.

Standard Curve

References:

1: Soliman MM, Taghi-Kilani R, Abou-Shady AF, El-Mageid SA, Handousa AA, Hegazi MM, Belosevic M.Comparison of serum antibody responses to Giardia lamblia of symptomatic and asymptomatic patients. Am J Trop Med Hyg. 1998 Feb;58(2):232-9.
2: Guimarães S, Sogayar MI. Detection of anti-Giardia lamblia serum antibody among children of day care centers. Rev Saude Publica. 2002 Feb;36(1):63-8.
3: Ljungström I, Castor B.Immune response to Giardia lamblia in a water-borne outbreak of giardiasis in Sweden. J Med Microbiol. 1992 May;36(5):347-52.
4: Wittner M, Maayan S, Farrer W, Tanowitz HB.Diagnosis of giardiasis by two methods. Immunofluorescence and enzyme-linked immunosorbent assay. Arch Pathol Lab Med. 1983 Oct;107(10):524-7.
5: Janoff EN, Smith PD, Blaser MJ.Acute antibody responses to Giardia lamblia are depressed in patients with AIDS. J Infect Dis. 1988 Apr;157(4):798-804.
6: Pérez O, Lastre M, Bandera F, Díaz M, Domenech I, Fagundo R, Torres D, Finlay C, Campa C, Sierra G. Evaluation of the immune response in symptomatic and asymptomatic human giardiasis. Arch Med Res. 1994 Summer;25(2):171-7.

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