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5-Hydroxyindoleacetic Acid (5-HIAA) ELISA Kit

INTENDED USE

5-HIAA ELISA kit is a procedure for accurately analysing in vitro quantitative levels of 5-hydroxyindoleacetic acid (5-HIAA) using urine samples. This assay has a minimum sensitivity detection limit of 0.17 mg/l.

BACKGROUND

5-hydroxyindoleacetic acid (5-HIAA) is in fact the main metabolite of serotonin and is often used to determine serotonin levels in the body by analysing urine samples of 5-HIAA. Both of these are produced in excess by many carcinoid tumors, in particular those forming carcinoid syndrome of hepatomegaly, flushing, bronchospasm, heart disease and diarrhea. This has led 5-HIAA being frequently being used for the diagnosis of carcinoid tumors of the enterochromaffin cells in the small intestines (this releases large quantities of serotonin). Elevated serotonin (also known as hyperserotonemia) is one of the primary biological findings in autism and there is evidence that 5-hydroxyindoleacetic acid may be elevated in patients with autistic spectrum disorders. Also, there have been cases of low levels of 5-HIAA found in the cerebrospinal fluid that have been associated with aggressive behaviour and suicide by violent means, there is a direct correlation with diminished serotonin levels.

This is a new procedure that is a simple method for the in vitro quantification of 5-hydroxyindoleacetic acid or 5-HIAA,  it is offering good sensitivity and producing accurate results for this important marker of carcinoid syndrome using only a small amount of urine sample. It has been well documented that a large number of substances that are present in urine have the ability to interfere with the traditional 5-hydroxyindoleacetic acid assayed by diazotization with nitrosonaphtol in order to form a purple colour. There have been many attempts to overcome this problem by a combination of fluorometry, ion exchange chromatography and even recently high performance liquid chromatography has been used. All of these methods are time consuming, require solvent extraction and lack sensitivity.

5-HIAA ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Adhesive Foil.
  • Reaction Tubes.
  • Reaction Plate.
  • Wash Buffer Concentrate (50x).
  • Enzyme Conjugate.
  • Substrate – Chromogenic substrate (TMB).
  • Stop Solution.
  • HIAA Microtiter Strips: Antigen pre-coated microwell plate.
  • Antiserum.
  • Standards (A-F) and Controls (1+2) – 0-50 mg/l.
  • Diluent.
  • Assay Buffer.
  • Methylation Buffer.
  • Methylation Reagent.

SENSITIVITY

The minimum sensitivity detection levels of 5-HIAA or 5-hydroxyindoleacetic acid using current 5-HIAA ELISA kit was approximately 0.17 mg/l. The dynamic range for this assay is 0.5 – 50 mg/l.

ASSAY CHARACTERISTICS

– 5-Hydroxyindoleacetic Acid (5-HIAA): ELISA
– Expected Reference Value: < 15 mg/day
– Measuring Range: 0.17 – 50 mg/l
– Analytical Sensitivity (Limit of Detection): 0.17 mg/l
– Analytical Specificity (Cross Reactivity): 5-HIAA (100%), Serotonin (5.5%), 5-Hydroxy-DL-Tryptophan (1.8%), Tryptamine (< 0.1%), Melatonin (< 0.1%), 5-Hydroxytryptamin (< 0.1%),  Vanillic – mandelic acid (< 0.1%),  Homovanillic Acid (< 0.1%).
– Intra Assay Precision: 8.6 – 14.1%
– Inter Assay Precision: 8.6 – 11.4%
– Linearity: 98 – 112%
– Recovery: 93 – 111%

REFERENCES

  1. Determination of urinary 5-hydroxyindoleacetic acid as a metabolomics in gastric cancer. J Gastrointest Cancer. (2015) 46 (2): 138-42. Mokhtari M., et al.
  2. The measurement of 5-hydroxyindoleacetic acid in urine. Ann Clin Biochem. (1994) 31 ( Pt 3): 215-32. Review. Deacon AC.
  3. The clinical and laboratory correlates of an increased urinary 5-hydroxyindoleacetic acid. Postgrad Med J. (1995) 71 (839): 542-5. Review. Tormey W.P. and FitzGerald R.J.
  4. Is urinary 5-hydroxyindoleacetic acid helpful for early diagnosis of acute appendicitis? Am J Emerg Med. (2012) 30 (4): 540-4. Jangjoo A., et al.

ADDITIONAL INFORMATION

  • Full Name: 5-Hydroxyindoleacetic Acid (5-HIAA) ELISA Kit
  • Reactivity: Any Species
  • Sample Type: Urine
  • Sensitivity:  0.17 mg/l

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