ANALYSIS OF SALIVARY ENZYME IN HUMAN

Saliva is an extracellular fluid secreted by salivary glands in the mouth. Saliva in humans, consists of water, electrolytes, mucus, white blood cells, epithelial cells, amylase, lipase, IgA, and lysozymes. Alpha amylase is present in humans, elephants, rats, and pigs. Amylase in humans is 42-59 U/ml.

STARCH IODINE PRESUMPTIVE TEST

Enzyme amylase is a component of saliva and can be detected by the addition of starchy solution followed by the addition of iodine. If amylase is not present in the sample, the iodine becomes trapped in the helical portions of starch forming starchy iodine complex (purple).If the amylase is present in the sample, the enzyme hydrolyzes starch to complex sugars and iodine cannot bind to the starch.  Amylase is also present in other biological materials such as feces, vaginal fluids, sweat and also in urine.

PHADEBAS® PRESUMPTIVE TEST

A tablet that is water-insoluble starch crosslinked to blue dye is hydrolyzed to blue fragments in the presence of alpha amylase.

RAPID STAIN IDENTIFICATION TEST(RSID) FOR SALIVA

RSID™ is a lateral flow immunochromatographic strip test for the detection of human saliva alpha amylase enzyme. Saliva uses two mouse monoclonal antibodies specific for salivary alpha amylase. One of the antibodies is conjugated to colloidal gold and is deposited on the conjugated pad. The other antibody is striped onto the ‘Test line’ on a membrane attached to the conjugate pad. Attached to the other end of the membrane is the wick, that absorbs the tested fluid and running. When the sample (tested fluid) is added to the sample window, the running buffer and sample diffuse through the conjugate pad, re-dissolving the gold-conjugated detection antibodies. If human salivary amylase is present in the sample an antigen-colloidal gold conjugated antibody complex will be formed producing a red line at the test and control position. If human salivary enzyme is not present in the sample a red line will appear at the control position on each strip.

 

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