Clinical Chemistry Exam 1 Lecture 5 – Flashcards
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Kjeldahl |
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Classic method for quantitating total protein |
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Kjeldahl = |
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Total Nitrogen Content x 16% |
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Biuret |
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Routine method for quantitating total protein |
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Biuret process |
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Cupric ions + peptide bonds = Violet Required 2 or more peptide bonds and an alkaline medium Color is proportional to protein concentration |
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For Biuret you should read color on a spec at ____ nm |
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540 |
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Should you avoid hemolysis with Biuret |
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Yes |
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Dye-Binding is often associated with which type of substance? |
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Albumin |
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Albumin + dye absorbs light at about _____ nm |
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600 |
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True or False: Absorbance is directly proportional to albumin concentration with Dye-Binding testing method |
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True |
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Dye Binding Principle |
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Positive charged Albumin + Negative charged dye |
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What types of dyes are associated with Dye-Binding? |
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Bromcresol green Bromcresol purple Methyl Orange |
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Nephelometry |
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Serum proteins added to specific antibody Detectors measure light scatter caused by ag-ab complexes Amount of light is proportional to the protein's concentration |
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What substances are often associated with Nephelometry |
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Tf Hapt Immunoglobulins C' |
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Is Serum Protein Eelectrophoresis quantitative or semi-quantitative? |
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Semi-quantitative |
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With the SPE in what direction will negatively charged proteins travel? |
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Toward the anode (+) |
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What does the rate of migration with the SPE depend on? |
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Size Charge Shape Time Support Media Endosmosis |
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What are the support media associated with SPE? |
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Cellulose acetate Agarose gel Polyacrylamide gel |
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Stains used with SPE? |
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Ponceau S Amido Black Coomassie Blue |
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A _____ converts visible protein bands into peaks. The area under each peak can be semi-quantitated. |
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Densitometer |
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Monoclonal gammopathy |
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[image] |
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Polyclonal gammopathy |
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[image] |
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Hypogammaglobulinemia |
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[image] |
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Alpha 1-Antitrypsin deficiency |
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[image] |
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Beta-Gamma bridging |
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[image] |
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Nephrotic Syndrome |
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[image] |
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Normal SPE |
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[image] |
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Proliferation of one clone of immunoglobulin |
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Monoclonal gammopathy |
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What diseases are associated with Monoclonal Gammopathy |
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Multiple Myeloma Waldenstrom's Macroglobulinemia |
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True or False: SPE does not identify the specific protein |
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True |
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What test would you run after SPE? |
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IFE |
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True or False: M spike may appear in alpha 2, beta, or gamma regions |
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True |
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What type of SPE are Osteolytic lesions and elevated calcium often associated with? |
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Monoclonal Gammopathy |
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Increase in all gamma proteins |
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Polyclonal gammopathy |
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What illnesses are often associated with Polyclonal Gammopathy |
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Chronic inflammation Infections |
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Hypogammaglobulinemia is associated with what type of illness? |
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Immunodeficiency |
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Is Hypogammaglobulinemia inherited or acquired? |
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Both |
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How can Hypogammaglobulinemia be acquired? |
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Cancer, chemo, HIV, etc. |
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What illness is associated with Alpha 1-Antitrypsin deficiency? |
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Emphysema |
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Beta-gamma bridging is associated with what type of gamma proteins? |
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Fast-moving |
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What illness is Beta-gamma bridging associated with? |
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Liver cirrhosis |
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In Nephrotic syndrome what substance is associated with increased protein loss through the kidneys? |
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Acute phase reactants |
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In Nephrotic syndrome what substance is associated with decreased protein loss through the kidneys? |
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Albumin |
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Globulin = |
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Total protein - Albumin |
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Normal Ratio of A/G |
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1.5 to 1 |
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Low A/G ratio |
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Too many globulins or Too little albumin |
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High A/G ratio |
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Not enough globulins |
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What illnesses are associated with: low A/G ratio due to too many globulins |
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MM Autoimmune diseases |
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What illnesses are associated with: low A/G ratio due to too little albumin |
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Renal disease Liver disease |
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What illnesses are associated with: not enough globulins |
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Genetic Leukemia Immunosuppressed |
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Immunofixation Electrophoresis |
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Identifies the specific protein causing the M spike on SPE or UPE |
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Normal Urine protein electrophoresis |
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Urine concentrated x 100 |
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Normal Urine protein electrophoresis shows only a _____ amount of albumin |
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small |
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Urine with Bence Jones Protein |
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Obvious M Spike IFE needed to identify type of light chain present |
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What type of band is more obvious with a Normal CSF Electrophoresis? |
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prealbumin |
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CSF with Oligoclonal Bands |
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Abnormal banding in the gamma region IgG Index >0.8 |
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What disease is CSF with Oligoclonal Bands consistent with? |
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Multiple Sclerosis |
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Which type of Electrophoresis is fast and requires little sample? |
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Capillary (Zone) Electrophoresis |
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With Capillary Electrophoresis, when the capillary is connected to a power supply, the charged molecules migrate at different rates depending on what? |
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net charge |
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Capillary Electrophoresis: ____ are measured by absorbance as they pass through a detector |
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Proteins |
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Plasma Viscosity |
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Non-specific measure of protein changes due to inflammation/tissue damage |
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Does Plasma Viscosity Increase or Decrease in parallel to the ESR |
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Increase |
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Is Plasma Viscosity affected by hematocrit or testing delays? |
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No |
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What diseases is an increased plasma viscosity associated with? |
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Myeloma Hyperfibrinogenemia |
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How is Plasma Viscosity measured with? |
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Viscometer |
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What are the reporting units of Plasma Viscosity? |
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Centipoises (cP or cps) |
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What is the Normal Rate of Plasma Viscosity? |
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1.1-1.5 cP |
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Water at Room Temperature is about ___ cP |
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1 |
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What is the Critical Value for Plasma Viscosity? |
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>3.0 cP |
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If G is increased or A is decreased the ratio will be low. What 3 diseases are associated with this? |
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MM Chronic Disease Nephrotic Syndrome |
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If G is decreased the ratio will be high. What disease is associated with this? |
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Immune Suppression |