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
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