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Clinical Biochemistry Flashcards

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1
Q

Define clinical biochemistry

A

Clinical analysis of bodily fluids for diagnosis, therapy and prevention of disease

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2
Q

What are typical panels measured in biochemistry?

A 
Fluids from serum, urine and joints
Liver- ALT, GLDH, ALP. GGT, bilirubin, bile acids
Kidney- urea, creatinine
Proteins- TP, albumin
Electrolytes- Na+, K+, Cl-, Ca2+, PO4-
Glucose and lipids
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3
Q

What quality control measures for biochemistry should be made?

A 
System set up
Maintenance
Cleaning
Storing samples
Interpretation of results
Control tests
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4
Q

What is plain tubes used for and some examples?

A

For samples that are allowed to clot and have serum separated
Bile acids, protein electrophoresis
Dont use for fibrinogen

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5
Q

What are heparin tubes used for and how do they work?

A

Plasma

Stops clot formation by increasing action of antithrombin III

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6
Q

What is OxF tube used for?

A

Measuring exact glucose levels

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7
Q

What are citrate tubes used for and how do they work?

A

Haemostasis

Anticoagulant binds to calcium

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8
Q

What are EDTA tubes used for and how do they work?

A

Haematology

Contains potassium and uses up calcium

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9
Q

What are examples of biochemistry analysers?

A

Glucometers
Wet/dry biochemistry analysers
Electrolyte analyser
Blood gas analyser

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10
Q

What are biological factors that affect results?

A

Interindividual- differences between groups of animals

Intraindividual- differences within one individual, should be minimised as much as possible

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11
Q

What pre analytical factors affect results?

A 
Poor sampling
Haemolysed, lipemic or icteric plasma
Wrong container
Wrong anticoagulant
Storage of sample
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12
Q

What is lipemia and what effects does it have on samples?

A

Lipids in serum causing milky with visible turbidity

Can dilute other substances and interferes with haematology and spectrophotometric assays

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13
Q

What is icterus in samples?

A

Jaundice/increased bilirubin so yellow serum

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14
Q

What is haemolysis of samples and what effects does it have on analysis?

A

Red plasma from free haemoglobin

Interferes with spectrophotometric assays, haematology and chemical interactions

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15
Q

How can haemolysis be prevented?

A

Good sampling technique
Avoid delays
Keep refrigerated
Separate serum

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16
Q

What are reference intervals?

A

Normal value for 90% of healthy population

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17
Q

What causes high levels of bilirubin?

A

RBC breakdown, liver disease, bile duct obstruction

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18
Q

What causes high levels of bile acids?

A

Decreased hepatic function and bile flow from portosystemic shunt

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19
Q

What liver enzymes increase with liver damage?

A

ALT
AST
GLDH
Cholestatic enzymes

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20
Q

What substances show liver function?

A 
Urea
Glucose
Albumin
Bile acids
Bilirubin
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21
Q

How to run bile acid stimulation test?

A

Take fasted sample
Feed
Collect sample 2 hours later

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22
Q

What do different pancreatic enzymes suggest is a cause?

A

Pancreatic lipase immunoreactivity- pancreatic injury

Trypsinogen like immunoreactivity- exocrine pancreatic functional mass

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23
Q

What is azotaemia?

A

Increased serum urea

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24
Q

What are the different types of azotaemia?

A

Prerenal- decreased renal perfusion, shock, CV disease, dehydration
Renal- renal disease
Post renal- urinary tract obstruction causes accumulation and rupture so waste enters abdomen

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25
Q

What are clinical causes of high blood glucose and most common causes of low blood glucose in samples?

A

High- stress, diabetes, steroids

Low- incorrect sample handling, insulinoma tumour, insulin overdose

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26
Q

What causes increased creatine kinase?

A

Skeletal muscle injury

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27
Q

What do urea and creatinine levels measure?

A

GFR

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28
Q

What causes changed levels of urea and creatinines?

A

Increased- dehydration, renal disease, urinary obstruction, heart disease
Decreased urea- liver failure
Decreased creatinine- muscle wasteage

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29
Q

What are the proteins known as total proteins and what are their purpose?

A

Albumin and globulins

Maintain colloid osmotic pressure

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30
Q

What are consequences of changed levels of total proteins?

A

Increase- lipemia (false), dehydration, inflammation

Decrease- haemorrhage, GI disease, renal disease, hepatic disease

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31
Q

What causes increased and decreased albumin?

A

Increased- dehydration

Decrease- inflammation, liver disease, kidney disease, GI disease, haemorrhage

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32
Q

How does serum protein electrophoresis work?

A

Elevates groups of protein in serum showing fraction of albumin and globulin

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33
Q

Define haemostasis

A

Ability to stop bleeding

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34
Q

What are the stages of haemostasis?

A

Primary- rapid unstable response, platelets form plug which helps activate other platelets
Secondary- fibrin mesh formation to stabilise platelet plug
Tertiary- break down of clot to return normal vascular flow by preventing overclotting

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35
Q

What are the consequences of haemostasis disorders?

A

Defective- haemorrhage

Excessive- thromboembolism

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36
Q

List lab tests can assess haemostasis

A 
Buccal mucosa bleeding time
Platelet numbers
Clotting time
Fibrinolysis tests
Platelet function tests
Genetic tests
Individual clotting factors
Thromoelastography
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37
Q

Explain the buccal mucosal bleeding time test and what it tests for

A

Cut MM
Collect blood with filter paper until bleeding stops
Normal time takes 3.3 minutes or less
Tests primary haemostasis

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38
Q

Explain how clotting times can be tested and what the normal values can be

A

Collect whole blood and allow to clot
Dogs- less than 90 seconds
Cats- less than 60 seconds

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39
Q

What are clinical disorders of haemostasis?

A

Primary or secondary- GI bleeding, epistaxis, haematuria

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40
Q

What are the most common causes of haemostasis disorders?

A

Low platelets

Low production, over use due to haemorrhage or increased destruction

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41
Q

Define thrombocytopenia

A

Low platelets

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42
Q

Define haematogram

A

Shows erythrocyte leukocyte and platelet parameters

Platelets should be double checked

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43
Q

How does haematology analyser generate parameters?

A

Laser- RBC count and mean cell volume
Lyses cells- haemoglobin
Calculated- haematocrit, mean corpuscular haemoglobin and concentration

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44
Q

How to store blood samples?

A

Fill EDTA tube to line
Invert and roll to prevent clotting
Store in fridge
Prepare and dry smears but dont put in fridge

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45
Q

Define haematocrit

A

Proportion of blood that is red blood cells

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46
Q

What are evaluations that can be made about RBC from a sample?

A

Circulating RBC mass- haematocrit, PCV, RBC count
What RBC looks like- mean corpuscular volume, haemoglobin and conc
RBC morphology- peripheral blood smear

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47
Q

Define PCV

A

Packed cell volume

Percentage of red blood cells in blood

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48
Q

How do you read PCV?

A

Centrifuge whole blood
Read as % of column
Shows plasma, buffy coat of WBC and platelets, packed red cells

49
Q

What are suffixes associated with haemostasis?

A

Philia/cytosis- increase in number
Penia- decrease in number
(Philia- granulocytes
Cytosis- all other cells)

50
Q

Describe flow cytometry in haematology

A

Individual cells pass through laser, absorbing and scattering light
Light interruptions count cells
Scattering determines size and complexity

51
Q

Describe impedance in haematology

A

Cells pass in isotonic solution between electrolytes and are poor electrical conductors
Change in electrical impedance is proportional to cell size

52
Q

What interferes with haematology analysers?

A 
Clots
Platelet clumps
RBC agglutination
Nucleated RBC
Lipemia
Leukocyte agglutination
Handling delays
53
Q

What are the different areas of a blood smear?

A

Base- not examined
Monolayer- main bulk with RBC not overlapping
Feathered edge

54
Q

How do you carry out a leukogram?

A

Identify cell types
Count number
Smear exam of monolayer

55
Q

Why do you need to do a blood smear?

A

Automated analysers dont pick up morphology changes

Help make quick clinical choices

56
Q

How to analyse a blood smear?

A

Low magnification at feathered edge
Move into monolayer
Increase to oil layer at monolayer
Move to lateral edge and count 100 leukocytes into types and observe abnormal forms

57
Q

What RBC observations can you make from blood smear?

A

Number
Normality
Evidence of regeneration

58
Q

What are normal morphology RBC in dogs and cats?

A

Dogs- uniform, pale central 1/3 of cell

Cats- small, uniform, no central colour

59
Q

What platelet observations can you make from blood smear?

A

Number

Morphology

60
Q

What do platelets look like in a blood smear?

A

Size of RBC
No nucleus
Granulation
Can be in clumps

61
Q

What observations can you make for WBC from blood smear?

A

Number
Type
Morphology

62
Q

What are different types of WBC and what do they look like?

A

Neutrophils- segmented ribbon shaped nucleus, pale cytoplasm
Eosinophils- pink granules
Basophils- purple granules
Lymphocytes- round with round nucleus and little cytoplasm
Monocytes- granular appearance, some have vacuoles

63
Q

How do you estimate platelet counts?

A

Count platelets in 10 fields using oil immersion lens in monolayer
Calculate average and multiply by 15 or 20 then x10^9

64
Q

What is the normal average number of platelets counted in monolayer?

A

15-30

65
Q

What are the problems of automated platelet counts?

A

Commonly lead to artefactual thrombocytopenia due to clumping or macroplatelets
RBC and platelets are same size so can be miscounted

66
Q

When should you do a blood smear for platelet counts?

A

Signs of haemorrhage

Low automated counts

67
Q

What is observed about RBC morphology for the different types of anaemia?

A

Non-regenerative- few normal looking RBCs

Regenerative- few abnormal looking RBCs

68
Q

What are causes of anaemia?

A

Reduced RBC count
Reduced haemoglobin concentration
Low PCV

69
Q

What are clinical signs of anaemia?

A 
Pale/yellow MMs
Lethargy
Tachycardia
Tachypnoea
Collapse
GI blood
Pica
70
Q

How can you classify anaemia?

A

RBC indexes
Regenerative or non-regenerative
Severity

71
Q

When does regeneration of RBC naturally take place and how is it determined?

A

Low oxygen

Reticulocyte concentration

72
Q

What are reticulocytes?

A

RBC precursors, 2 types

73
Q

What are the types of reticulocytes?

A

Aggregate- get counted, immature, released by bone marrow in response to anaemia, have lots of dots in cytoplasm
Punctate- matured, stay in blood so unreflective of current status, counted as % in blood smear

74
Q

What are types of anaemia when based off mean cell volume?

A

Normocytic- normal sized erythrocytes
Microcytic- low MCV, iron deficiency causes division into small RBCs
Macrocytic- high MCV, immature RBC in circulation larger than mature RBCs

75
Q

What are types of anaemia based off haemoglobin levels?

A

Normochromic
Hypochromic- iron deficiency, immature RBCs
Hyperchromic- usually are arterfact

76
Q

What are common morphological changes of RBCs?

A

Anisocytosis- different cell sizes
Polychromasia- indicate regeneration, RBC look purple
Hypochromasia- low haemoglobin so larger pale area
Spherophytes- small, round, dark
Ghost cells- only membrane of RBC present

77
Q

What are signs of thrombocytopenia?

A

Petechiae/pinpoint haemorrhage
Bruises
GI bleeding
Spontaneous haemorrhage

78
Q

How is thrombocytopenia diagnosed?

A

Blood smear

Rerun haematology

79
Q

Define polycythaemia and what is its causes

A

Increased red cell mass due to increased haemoglobin, PCV

80
Q

What are the types of polycythaemia?

A

Erythrocytosis- relative due to plasma loss/dehydration

True polycythaemia- actual increased mass

81
Q

What are signs of polycythaemia?

A 
High PCV
Dark MM
Sneezing
Nose bleeds
Neurological signs
82
Q

How is polycythaemia diagnosed?

A 
Check hydration
Lab tests
Blood gas
Ultrasound
Radiography
83
Q

What are common changes causing WBC disorders?

A 
Stress
Inflammation
Adrenaline
Neoplasia
Inverted stress
84
Q

Define neutrophilia

A

Increased neutrophils due to infection

85
Q

Define neutropenia

A

Decrease of neutrophils due to bone marrow destruction or suppression

86
Q

How do you count WBCs from blood smears?

A

Take at edge of monolayer and move up and down

Count percentage of types of leukocyte

87
Q

What are the morphologies of lymphocytes?

A

Normal- round, double size of RBC, little cytoplasm
Reactive- smaller nucleus and larger cytoplasm
Lymphoblast- large nucleus

88
Q

Define lymphocytosis and when is it seen?

A

High lymphocyte counts
Normal in young for immune development
Epinephrine release, cell mobilisation, antigenic stimulation, cancer

89
Q

What is monocytosis and when is it seen?

A

Increased demand and production of monocytes

Infection, inflammation and trauma

90
Q

What is eosinophilia and what are causes?

A

Increased demand of eosinophils

Allergy, inflammation, parasites, eosinophil leukaemia

91
Q

What is lymphopenia and what are caused?

A

Loss of or decreased production of lymphocytes

Virus or immunodeficiency

92
Q

Define urinalysis

A

Physical, chemical and microscopic analysis of urine to diagnose and manage disease

93
Q

What are benefits of urinalysis?

A

Rapid in house test, transport effects sample

Cheap and basic equipment

94
Q

What are the stages of urinalysis?

A

Physical exam
Chemical analysis
Sediment analysis
Uroculture

95
Q

What equipment is needed for urinalysis?

A 
Urinary strips
Refractometer
Centrifuge
Tubes
Microscope
Sediment stain
Slides
Cytological stain
96
Q

How can you collect urine for sampling?

A 
Off floor
Free catch
Bladder squeeze
Catheterisation
Cystocentesis
97
Q

How to store urine samples, including the tubes used for free catch and cytology samples?

A

Free catch- boric acid tube to prevent growth of contaminant bacteria
Cytology- EDTA tube
Store in fridge if analysis takes longer than 30 minutes, but does cause crystal precipitation
Avoid direct sunlight
Return to room temperature before analysis

98
Q

Describe the process of sediment analysis

A

Mix well before analysis
Centrifuge 5mls
Decant supernatant leaving 0.5ml for resuspension
Stain when needed
Resuspend until well mixed
Add to slide and examine under microscope

99
Q

What should be seen under a microscope in urine?

A

Less than 5 erythrocytes and leukocytes per 500x field
Squamous epithelial cells in free catch, large flat cells
Transitional epithelial cells, large cells
Crystals

100
Q

What are different urine crystals?

A 
Struvite- rooftop shaped
Calcium oxalate- square with cross
Bilirubin- branched
Biurate- round with spikes
Calcium carbonate
101
Q

What gross appearance should be noted in urine samples?

A

Turbidity
Colour
Odour

102
Q

How do you measure specific gravity of urine?

A

Refractometer, quality control with water

Measure at room temperature

103
Q

What are the interpretations of urine specific gravity?

A

Hypersthenuria- concentrated urine, normally hydrated animals
Isosthenuria- not concentrated or dilute, investigate when persistent
Hyposthenuria- diluter than plasma, investigate when persistent

104
Q

What parameters are measured on urine dipstick tests?

A

Reliable- pH, protein, ketones, bilirubin, blood

Unreliable- specific gravity, urobilinogen, nitrate leukocytes

105
Q

What effect does storage have on pH of urine?

A

Rapidly makes more alkaline

106
Q

What needs investigating regarding protein levels in urine?

A

Proteinuria in dilute samples or >1 when concentrated sample

107
Q

What are causes of proteinuria (pre- post- and renal)?

A

Pre-renal- hyperproteinaemia/high protein in the blood, hyperthermia, seizures, venous congestion
Renal- glomerular, tubular
Post-renal- inflammation, haematuria

108
Q

What causes false negative bilirubin and what causes high bilirubin?

A

False negatives- light

High- liver disease, haemolytic anaemia

109
Q

What causes ketones to be present in the urine?

A

Diabetic ketoacidosis
Ketosis
Starvation

110
Q

What are causes of glucose in the urine?

A

Hyperglycaemia
Renal tubular disorders
False positives- bleach, hydrogen peroxide

111
Q

What does nitrate in the urine suggest?

A

UTI

112
Q

What are the benefits of in house labs?

A 
Fast turn around
Rapid treatment
Monitoring
Smaller volumes required
Cheaper after initial investment
113
Q

What are post regulations of sending samples?

A 
50ml limit
Sealed container
Padding enough to absorb entire contents
Leak proof bag
No class 4 pathogens
114
Q

What needs to be included when sending samples to external lab?

A

ID of patient, owner and vet practice
Sample type
Tests required
Any history

115
Q

How to avoid haemolysis when collecting blood samples?

A

Clean stick

No vacuum created

116
Q

What are consequences of stressful samples?

A 
Neutrophilia
Lymphocytosis
Elevated PCV
Elevated creatine Kinase with restraint
Haemolysis
117
Q

How do serum samples differ to plasma samples?

A

Serum- long time to clot, need spinning later

Plasma- need anticoagulant, spin and separate immediately

118
Q

What happens if EDTA tubes get contaminated?

A

Alters electrolyte levels and some enzymes

119
Q

What is the best collection for cytology?

A

Fine needle aspirate biopsies

Fluids in EDTA and plain tube for culturing

Pre-Clinical Science (5 decks)

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