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Chemistry year 2 > Week 11 > Flashcards

Week 11 Flashcards

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1
Q

3 major classes of plasma proteins

A

albumin, globulins and fibrinogen

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2
Q

Function of albumin

low levels are seen in

testing usually done with

A

maintains Collodial osmotic pressure
regulates fluid balance
most abundant protein in blood

liver disease, nephrotic syndrome, malnutrition, or chronic inflammation

tested with bromocresol green or purple DYE BINDING METHOD - calibrate and use controls- avoid hemolysis

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3
Q

Function of Fibrinogen

A

protein in blood clotting
converts to fibrin to form clot

HIGH levels in ACUTE inflammation, pregnancy, and birth control use

LOW levels in liver disease, DIC, and malnutrition

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4
Q

how do you know if you have nephrotic syndrome

A

swelling because you have low protein = low plasma proteins
albumin is lost through urine

proteinuria- frothy urine - DIPSTICK
hypoalbuminemia - muscle wasting - tiredness HALLMARK
hyperlipidemia - lipid in urine
edema (periorbital swelling and anasarca
glomerulus injury -proteins leak into urine
renal biopsy

macroscopic testing detects lipid casts and oval fat bodies

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5
Q

how to do you know you have liver disease

A

altered albumin and globulin levels will show you have -cirrhosis- alchohol, Hep B/C, NAFLD
-hepatitis-inflamed liver due to virus A-E
-fatty liver disease- insulin resistance, AFLD from alcohol
-Liver cancer - hepatocellular carcinoma HCC - primary liver cancer

symptoms
juandice
ascites - fluid in abdominal cavity
hepatomegatly
coagulopathy

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6
Q

What will you see in Chronic Inflammatory and Autoimmune Disorders

A

-increased gamma globulins because your immune system is constantly stimulated

-protein electrophoresis can help to differentiate between polyclonal and monoclonal

RA, SLE and IBD

increase of cytokines TNF alpha
autoantibody production

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7
Q

What quality things can affect plasma protein testing

A

Sample quality- hemolysis, lipemic and icteric samples

Calibration and controls - regular calibration and QC with EVERY BATCH

check to make sure there are no interferences like medications or paraproteins

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8
Q

What is CRP

A

initatites opsonization and encourages phagocytosis
-marker of early phase reactions and increases with inflammation, bacterial and viral infections, MI, RA, gout

increased when there is liver disease, malnutrition and protein losiing

risk factor for CVD, inflammation and atherosclerosis
-part of non specific response
-increases quickly in 6-12 hours of inflammation and does down once the inflammation goes down

-flareups associated with IBD or SLE

hs CRP = increased with atherosclerosis, coronary artery disease, and atherosclerosis

monitored post op to detect infections

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9
Q

IgG associated with

A

autoimmune disease and hepatic disease

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10
Q

IgA associated with

A

skin, gut, respiratory infections, cirrhosis

found in secretions like tears

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11
Q

IgM associated with

A

viral infections, blood infections (malaria), and acute hepatitis.

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12
Q

IgD associated with

A

surface receptor for antigens on B lymphocytes
primary function unknown

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13
Q

IgE associated with

A

histamine release from mast cells
asthma, allergies and parasites

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14
Q

what are poly and monoclonal increases in gammaglobulins associated with

decreases in gammaglobulins associated with

A

poly increases = chronic infections, liver disease, autoimmune and parasite infections

mono increases - myeloma , Waldenström
macroglobulinemia

decreases = leaves you vulnerable to diseases and can order through primary or secondary causes like sickness or drugs

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15
Q

What is the albumin/globulin ratio

A

-gives you albumin ratio by dividing up the globulin concentration
-RI is 1.0-1.8 variations can occur because of lab methods
-looks at liver function, immune function

Globulins = TP - ALB

then A/G ratio is A/G

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16
Q

What does a low A/G ratio tell you

A

less than 1.0

-increased globulins and decreased albumin

CHRONIC liver disease - reduced protein syn
Nephrotic syndrome - albumin lost through urine because of kidney damage

Chronic inflammatory or autoimmune disorders- RA, Lupus = high globulins

Myeloma, plasma cell dyscrasias - over production of gammaglobulins

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17
Q

what does a high A/G ratio tell you

A

high than 2.5

Hypogammaglobulinemia- Common Variable Immunodeficiency - CVID

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18
Q

what does increased vs decreased Total protein tell you

A

myeloma, chronic infections, and dehydration.

decreased:
nephrotic syndrome, severe burns, liver disease,
inflammation, and malnutrition.

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19
Q

how are protein levels measured

A

Spectrophotometry, immunoassay, nephelometry, electrophoresis, and mass spectrometry

20
Q

What is the Biruet method of measuring total protein

A

color change that occurs when cupric ions are in alkaline solution
-color intensity is proportional to protein concentration looked at through spectrophotometer

-hemolysis, icterus , lipemia can cause issues with measurement

-contains
CuSO4, NaOH, NaK tartrate, KI.

21
Q

How is albumin measured

A

bye binding
Nephelometry, turbidimetry for specific proteins.

22
Q

What do abnormal SPE show you

A

patterns for nephrotic syndrome and acute phase proteins.
Abnormal SPE in Various Conditions

23
Q

how is liver cirrhosis seen on SPE

A

Increased β-γ globulins indicative of cirrhosis.

24
Q

how is Hypogammaglobulinemia seen on SPE

A

Decreased or absent γ-globulins, suggesting immune deficiencies.

25
Q

how is Monoclonal Gammopathy seen on SPE

A

M-spike in γ-globulin region, common in multiple myeloma., Waldenström’s macroglobulinemia, and MGUS

26
Q

What other testing indications will you find in multiple myeloma

A

bence jones protein = kappa or lamba free light chains

increased IgG, hypercalcemia, renal insufficiency, anemia and bone lesions

27
Q

What is Waldenstrom Macroglobulinemia

A

non-Hodgkin’s lymphoma
severe fatigue , bleeding and anemia

can use IFE to detect = IgM monoclonal proteins

28
Q

how does IFE work and what are its problems

A

two stage procedure
separate proteins according to motility and immunoprecipitated to detect abnormal Immunoglobulins

used for monoclonal proteins looking for MS

if there is AG excess , artifacts, improper washing it can cause inaccuracies in the IFE results

29
Q

How can refractometery be used to measure proteins

A

uses the refractive index of serum which changes with the protein concentration in serum
-less accurate/specific than biuret method

-is affected by glucose and electrolytes in sample

30
Q

What is an interference in the dye binding method

A

high amounts of globulins, bilirubin = over estimation of albumin levels

31
Q

What is the immunoassay method for albumin detection

A

use antibodies specific to albumin to detect and
quantify its concentration.
AG-AB complexes are measured using nephelometry, turbidimetry

more costly

32
Q

What is SPE

A

separates and quantifies proteins in serum based on size, shape and charge

proteins separate into distinct bands: albumin, alpha-1, alpha-2, beta, and gamma globulins.

33
Q

What is the gel in Agarose gel electrophoresis stained with

A

protein-specific dye (e.g., Coomassie Brilliant Blue), and a densitometer is used to quantify the bands corresponding to different protein fractions (albumin, alpha-1, alpha-2, beta, and gamma).

34
Q

a broad increase in gamma region of electrohoresis gel tells you

A

its a poly clonal increases

35
Q

interferences in agarose gel electrophoresis

A

Hemolysis (free hemoglobin), lipemia (high levels of lipids), and paraprotein precipitation can cause artifacts that affect the interpretation of results.

36
Q

What is Capillary Electrophoresis

A

serum proteins are seperated in capillary tubes based on mobility in an electric field
-faster and better resolution
-uses an electrophoretogram to quantify protein fractions and detect monoclonal bands

37
Q

What is immunofixation electrophoresis IFE

A

detects and characterizes monoclonal proteins (also known as M-proteins or paraproteins) in serum,
urine, or cerebrospinal fluid (CSF).

used after SPE when a monoclonal protein is detected

-used to differentiate hematologic malignancies

38
Q

What is the gold standard for testing for multiple myloma

A

IFE
determines the heavy and light chain

39
Q

What is MGUS

A

Monoclonal Gammopathy of Undetermined Significance

detected by IFE
looking for monoclonal proteins in asymp proteins to monitor progression of multiple myeloma

40
Q

What is the procedure for IFE

A

first stage is like SPE where the proteins separate into distinct bands: albumin, alpha-1, alpha-2, beta, and
gamma globulins on gel with electric current

then antisera is applied to different gel sections to detect immunoglobulin heavy chains (IgG, IgA, IgM, IgD, IgE) and light chains (kappa and lambda).
-antisera binds to target proteins and forms immune complexes = fixed on get

-gel is then washed to removed anything that is unbound
-gel is stained with protein specific dyes like Coomassie blue or Amido Black , destained to remove excess dye

look at bands under imaging system

41
Q

what are common concerns for IFE troubleshooting

A

sample handling - make sure there is not hemolyis, lipemia or contamination

faint/absent bands - due to insufficient concentration improper antisera- REPEAT TEST WITH HIGHER SAMPLE CONCENTRATION

non specific binding - due to excessive background, wash carefully

distorted or diffuse bands - uneven polymerization , sample overloading

42
Q

how is CRP measured

A

immunoturbidimetry - changes in turbidity when CRP binds to ANTI crp antibodies
-measured with spectrophotometer and calculated based on calibration curve

-lipemia, hemolysis or icterus can impact test

43
Q

how is High-Sensitivity CRP (hs-CRP) Testing done

A

Immunonephelometry:
measuring light scatter by CRP AB complexes = detects low levels of CRP which is necessary for CVD
-concentration related to Calibration curve
-sensitive can detect down to 0.1mg/L = good for detection of low grade inflammation

hemolysis and lipemia can affect
<1 = low cardiovascular risk
>3 high risk

44
Q

How can ELISA be used to measure CRP

A

-binding CRP to surface bound AB followed by a 2ndary AB linked to enzyme , then a substrate which that is changed by the enzyme to a detectable signal

has a microplate coated with anti CRP antibodies

good for use in research
-but very labor intensive

45
Q

how can POCT be used for CRP testing

A

lateral flow immunoassay

capillary or venous
easy to use
less precise than lab test
hemolysis, lipemia and icterus is an issue
-different methods are best for specificity levels

46
Q

What tests would you order if you think someone had liver disease

A

ALT/AST - increased = hepatocellular injury
ALT = liver specific
AST = heart and muscles

ALP/GGT - increased in cholestasis and bile duct obstruction

Bilirubin - increased = liver dysfunction

ALB =decreased

PT - prolonged

Viruses - check for hepatitis

ANA, ASMA, Anti LKM

Imaging - Ultrasound, CT and MRI

47
Q

What tests to order if you think someone has Chronic Inflammatory and Autoimmune Disorders

A

Autoantibody tests: RF and ACPA = RA
ANA = SLE
anti ds DNA and Anti SM - very specific for SLE
ANCA= vasculitides or granulotomas

inflammatory markers - CRP, ESR
CBC
MRI and ultrasound for RA and arthritis
endoscopy for IBD

Chemistry year 2 (12 decks)

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