Term
| How much saliva do the human salivary glands produce per day? |
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Definition
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Term
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Definition
| A slightly alkaline secretion, comprised of water (mostly), mucus, proteins, salts, and enzymes |
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Term
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Definition
NO test is specific for saliva
Idntification of amylase indicates saliva |
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Term
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Definition
Enzyme that cleaves polysaccharidse that are composed of D-glucose units at the alpha1-4 linkages. AKA cleaves starch.
Digests carbohydrates in the oral cavity |
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Term
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Definition
| Contains two glucose polymers: amylose and amylopectin |
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Term
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Definition
| Long, unbranced glucose connected at the alpha1-4 |
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Term
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Definition
| Branched alpha1-6 connections and linear alpha1-4 |
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Term
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Definition
beta-amylase in plant and bacterial sources
alpha-amylase in human, human saliva (HSA), and human pancreatic (HPA) |
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Term
| Human alpha-amylases: Two isoforms |
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Definition
Differ in amino acid sequence
Human alpha-amylsase (HSA)
Human pancreatic amylase (HPA) |
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Term
| Human Alpha-Amylase (HSA) |
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Definition
From saliva
Inactivated by stomach acids
AMY1 locus |
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Term
| Human Pancreatic Alpha-Amylase (HPA) |
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Definition
From pancreas
Inactivated in the intestines with some activiy remaining in the feces.
AMY2 locus |
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Term
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Definition
| Semen, tears, milk, persperation, vaginal secretions, serum, and urine |
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Term
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Definition
Visual examination with ALS or microscope
Starch-iodine assays
Colorimetric Assays |
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Term
| Visual Examination: ALS or Microscope |
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Definition
Can see mucous cells to test if it is saliva
ALS: Fluorescence of saliva is less intense than that of semen
Microscopic: stain for buccal epithelial cells |
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Term
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Definition
Iodine used to test for starch
NOT specific because it is technically for STARCH
Starch appears blue, amylopectin appears reddish-purple in the presence of iodine
Albumin and other proteins produce a false positive |
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Term
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Definition
Dye-labeled amylase substrates are employed
Phadebas Test
Press Test |
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Term
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Definition
Clinical diagnosis & forensic use
Procion Red Amylopectin (PRA) or phadebas reagent:
-starch is linked to the dye forming insoluble complex
-dye becomes sluble, creating a colored product measured by a spectrophotometer
-done via paper or tube method
-may not detect low levels |
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Term
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Definition
-filters are sprayed with Phadebas solution and allowed to air dry
-paper soaked with dried Phadebas solution is pressed over the area to be tested
-initial time, single minutes for the first 10 minutes, and 5 minutes for up to 40 minutes total = intervals to note observations
Appears light blue on surface of paper |
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Term
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Definition
Immunochromatographic Assays - RSID-Saliva
ELISA - monoclonal HSA antibody
RNA-based Assays
-based on certain genes specifically expressed in certain types of cells
-based on mRNA & htRNA |
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Term
| Confirmatory Assays: Immunochromatographic Assays |
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Definition
RSID-Saliva
Sensititivy: 1 microL saliva necessary
specificity: no cross reaction with animal saliva, earwax, etc. most body fluids are okay.
utilizes 2 anti-salivary alhpa-amylase monocolonal antibodies in a lateral flow immunochromatographic strip test formation to detect the presence of salivary amylase rather than the activitiy of the enzyme
No high dose Hook effect |
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Term
| Confirmatory Assays: ELISA |
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Definition
monoclonal HSA (human salivary alpha-amylase) antibody
Only cross-reactivity for chimpanzee, mouse, rat, and eel
Did not work on old stains
Specific and can be used for quantification |
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Term
| Confirmatory Assays: RNA-based Assays |
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Definition
mRNA that is expressed only in the oral cavity
Uses reverse transcriptase - PCR (RT-PCR) to detect levels of mRNA
HTN3 encodes histain 3 - a protein involved in non-immune response host defense
STATH encodes statherin - inhibits precipitation of calcium salts in the salivary glands & oral cavity
Highly specific and can be automated, BUT RNA is unstable due to degradation |
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