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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
What are clinical causes of high blood glucose and most common causes of low blood glucose in samples?
High- stress, diabetes, steroids | Low- incorrect sample handling, insulinoma tumour, insulin overdose
26
What causes increased creatine kinase?
Skeletal muscle injury
27
What do urea and creatinine levels measure?
GFR
28
What causes changed levels of urea and creatinines?
Increased- dehydration, renal disease, urinary obstruction, heart disease Decreased urea- liver failure Decreased creatinine- muscle wasteage
29
What are the proteins known as total proteins and what are their purpose?
Albumin and globulins | Maintain colloid osmotic pressure
30
What are consequences of changed levels of total proteins?
Increase- lipemia (false), dehydration, inflammation | Decrease- haemorrhage, GI disease, renal disease, hepatic disease
31
What causes increased and decreased albumin?
Increased- dehydration | Decrease- inflammation, liver disease, kidney disease, GI disease, haemorrhage
32
How does serum protein electrophoresis work?
Elevates groups of protein in serum showing fraction of albumin and globulin
33
Define haemostasis
Ability to stop bleeding
34
What are the stages of haemostasis?
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
35
What are the consequences of haemostasis disorders?
Defective- haemorrhage | Excessive- thromboembolism
36
List lab tests can assess haemostasis
``` Buccal mucosa bleeding time Platelet numbers Clotting time Fibrinolysis tests Platelet function tests Genetic tests Individual clotting factors Thromoelastography ```
37
Explain the buccal mucosal bleeding time test and what it tests for
Cut MM Collect blood with filter paper until bleeding stops Normal time takes 3.3 minutes or less Tests primary haemostasis
38
Explain how clotting times can be tested and what the normal values can be
Collect whole blood and allow to clot Dogs- less than 90 seconds Cats- less than 60 seconds
39
What are clinical disorders of haemostasis?
Primary or secondary- GI bleeding, epistaxis, haematuria
40
What are the most common causes of haemostasis disorders?
Low platelets | Low production, over use due to haemorrhage or increased destruction
41
Define thrombocytopenia
Low platelets
42
Define haematogram
Shows erythrocyte leukocyte and platelet parameters | Platelets should be double checked
43
How does haematology analyser generate parameters?
Laser- RBC count and mean cell volume Lyses cells- haemoglobin Calculated- haematocrit, mean corpuscular haemoglobin and concentration
44
How to store blood samples?
Fill EDTA tube to line Invert and roll to prevent clotting Store in fridge Prepare and dry smears but dont put in fridge
45
Define haematocrit
Proportion of blood that is red blood cells
46
What are evaluations that can be made about RBC from a sample?
Circulating RBC mass- haematocrit, PCV, RBC count What RBC looks like- mean corpuscular volume, haemoglobin and conc RBC morphology- peripheral blood smear
47
Define PCV
Packed cell volume | Percentage of red blood cells in blood
48
How do you read PCV?
Centrifuge whole blood Read as % of column Shows plasma, buffy coat of WBC and platelets, packed red cells
49
What are suffixes associated with haemostasis?
Philia/cytosis- increase in number Penia- decrease in number (Philia- granulocytes Cytosis- all other cells)
50
Describe flow cytometry in haematology
Individual cells pass through laser, absorbing and scattering light Light interruptions count cells Scattering determines size and complexity
51
Describe impedance in haematology
Cells pass in isotonic solution between electrolytes and are poor electrical conductors Change in electrical impedance is proportional to cell size
52
What interferes with haematology analysers?
``` Clots Platelet clumps RBC agglutination Nucleated RBC Lipemia Leukocyte agglutination Handling delays ```
53
What are the different areas of a blood smear?
Base- not examined Monolayer- main bulk with RBC not overlapping Feathered edge
54
How do you carry out a leukogram?
Identify cell types Count number Smear exam of monolayer
55
Why do you need to do a blood smear?
Automated analysers dont pick up morphology changes | Help make quick clinical choices
56
How to analyse a blood smear?
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
What RBC observations can you make from blood smear?
Number Normality Evidence of regeneration
58
What are normal morphology RBC in dogs and cats?
Dogs- uniform, pale central 1/3 of cell | Cats- small, uniform, no central colour
59
What platelet observations can you make from blood smear?
Number | Morphology
60
What do platelets look like in a blood smear?
Size of RBC No nucleus Granulation Can be in clumps
61
What observations can you make for WBC from blood smear?
Number Type Morphology
62
What are different types of WBC and what do they look like?
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
How do you estimate platelet counts?
Count platelets in 10 fields using oil immersion lens in monolayer Calculate average and multiply by 15 or 20 then x10^9
64
What is the normal average number of platelets counted in monolayer?
15-30
65
What are the problems of automated platelet counts?
Commonly lead to artefactual thrombocytopenia due to clumping or macroplatelets RBC and platelets are same size so can be miscounted
66
When should you do a blood smear for platelet counts?
Signs of haemorrhage | Low automated counts
67
What is observed about RBC morphology for the different types of anaemia?
Non-regenerative- few normal looking RBCs | Regenerative- few abnormal looking RBCs
68
What are causes of anaemia?
Reduced RBC count Reduced haemoglobin concentration Low PCV
69
What are clinical signs of anaemia?
``` Pale/yellow MMs Lethargy Tachycardia Tachypnoea Collapse GI blood Pica ```
70
How can you classify anaemia?
RBC indexes Regenerative or non-regenerative Severity
71
When does regeneration of RBC naturally take place and how is it determined?
Low oxygen | Reticulocyte concentration
72
What are reticulocytes?
RBC precursors, 2 types
73
What are the types of reticulocytes?
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
What are types of anaemia when based off mean cell volume?
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
What are types of anaemia based off haemoglobin levels?
Normochromic Hypochromic- iron deficiency, immature RBCs Hyperchromic- usually are arterfact
76
What are common morphological changes of RBCs?
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
What are signs of thrombocytopenia?
Petechiae/pinpoint haemorrhage Bruises GI bleeding Spontaneous haemorrhage
78
How is thrombocytopenia diagnosed?
Blood smear | Rerun haematology
79
Define polycythaemia and what is its causes
Increased red cell mass due to increased haemoglobin, PCV
80
What are the types of polycythaemia?
Erythrocytosis- relative due to plasma loss/dehydration | True polycythaemia- actual increased mass
81
What are signs of polycythaemia?
``` High PCV Dark MM Sneezing Nose bleeds Neurological signs ```
82
How is polycythaemia diagnosed?
``` Check hydration Lab tests Blood gas Ultrasound Radiography ```
83
What are common changes causing WBC disorders?
``` Stress Inflammation Adrenaline Neoplasia Inverted stress ```
84
Define neutrophilia
Increased neutrophils due to infection
85
Define neutropenia
Decrease of neutrophils due to bone marrow destruction or suppression
86
How do you count WBCs from blood smears?
Take at edge of monolayer and move up and down | Count percentage of types of leukocyte
87
What are the morphologies of lymphocytes?
Normal- round, double size of RBC, little cytoplasm Reactive- smaller nucleus and larger cytoplasm Lymphoblast- large nucleus
88
Define lymphocytosis and when is it seen?
High lymphocyte counts Normal in young for immune development Epinephrine release, cell mobilisation, antigenic stimulation, cancer
89
What is monocytosis and when is it seen?
Increased demand and production of monocytes | Infection, inflammation and trauma
90
What is eosinophilia and what are causes?
Increased demand of eosinophils | Allergy, inflammation, parasites, eosinophil leukaemia
91
What is lymphopenia and what are caused?
Loss of or decreased production of lymphocytes | Virus or immunodeficiency
92
Define urinalysis
Physical, chemical and microscopic analysis of urine to diagnose and manage disease
93
What are benefits of urinalysis?
Rapid in house test, transport effects sample | Cheap and basic equipment
94
What are the stages of urinalysis?
Physical exam Chemical analysis Sediment analysis Uroculture
95
What equipment is needed for urinalysis?
``` Urinary strips Refractometer Centrifuge Tubes Microscope Sediment stain Slides Cytological stain ```
96
How can you collect urine for sampling?
``` Off floor Free catch Bladder squeeze Catheterisation Cystocentesis ```
97
How to store urine samples, including the tubes used for free catch and cytology samples?
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
Describe the process of sediment analysis
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
What should be seen under a microscope in urine?
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
What are different urine crystals?
``` Struvite- rooftop shaped Calcium oxalate- square with cross Bilirubin- branched Biurate- round with spikes Calcium carbonate ```
101
What gross appearance should be noted in urine samples?
Turbidity Colour Odour
102
How do you measure specific gravity of urine?
Refractometer, quality control with water | Measure at room temperature
103
What are the interpretations of urine specific gravity?
Hypersthenuria- concentrated urine, normally hydrated animals Isosthenuria- not concentrated or dilute, investigate when persistent Hyposthenuria- diluter than plasma, investigate when persistent
104
What parameters are measured on urine dipstick tests?
Reliable- pH, protein, ketones, bilirubin, blood | Unreliable- specific gravity, urobilinogen, nitrate leukocytes
105
What effect does storage have on pH of urine?
Rapidly makes more alkaline
106
What needs investigating regarding protein levels in urine?
Proteinuria in dilute samples or >1 when concentrated sample
107
What are causes of proteinuria (pre- post- and renal)?
Pre-renal- hyperproteinaemia/high protein in the blood, hyperthermia, seizures, venous congestion Renal- glomerular, tubular Post-renal- inflammation, haematuria
108
What causes false negative bilirubin and what causes high bilirubin?
False negatives- light | High- liver disease, haemolytic anaemia
109
What causes ketones to be present in the urine?
Diabetic ketoacidosis Ketosis Starvation
110
What are causes of glucose in the urine?
Hyperglycaemia Renal tubular disorders False positives- bleach, hydrogen peroxide
111
What does nitrate in the urine suggest?
UTI
112
What are the benefits of in house labs?
``` Fast turn around Rapid treatment Monitoring Smaller volumes required Cheaper after initial investment ```
113
What are post regulations of sending samples?
``` 50ml limit Sealed container Padding enough to absorb entire contents Leak proof bag No class 4 pathogens ```
114
What needs to be included when sending samples to external lab?
ID of patient, owner and vet practice Sample type Tests required Any history
115
How to avoid haemolysis when collecting blood samples?
Clean stick | No vacuum created
116
What are consequences of stressful samples?
``` Neutrophilia Lymphocytosis Elevated PCV Elevated creatine Kinase with restraint Haemolysis ```
117
How do serum samples differ to plasma samples?
Serum- long time to clot, need spinning later | Plasma- need anticoagulant, spin and separate immediately
118
What happens if EDTA tubes get contaminated?
Alters electrolyte levels and some enzymes
119
What is the best collection for cytology?
Fine needle aspirate biopsies | Fluids in EDTA and plain tube for culturing

Pre-Clinical Science (5 decks)

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