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Clinical Biochemistry Flashcards

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1
Q

define clinical biochemistry

A

clinical analysis of body fluids for the diagnosis, therapy and prevention of diseases

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2
Q

what can be sampled for biochemistry?

A

usually serum/plasma but can be urine and other body fluids (e.g. pleural fluid)

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3
Q

what body systems/parameters does a typical biochemistry panel include?

A 
liver 
kidney
proteins
electrolytes 
glucose
lipids
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4
Q

what are panels separated into?

A

organ or system

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5
Q

what liver parameters are typically assessed on a biochemistry panel?

A

ALT, AST, GLDH, ALP, GGT, Bilirubin, bile acids

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6
Q

what liver parameters demonstrate liver damage?

A

hepatocellular damage/ leakage enzymes - ALT, AST, GLDH

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7
Q

what level of liver damage does ALT show?

A

mild

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8
Q

what level of liver damage does AST show?

A

necrosis

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9
Q

what liver parameters demonstrate bile accumulation?

A

cholestatic enzymes - ALP, GGT

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10
Q

what substances can be measured on a biochemistry panel that are conjugated and excreted by the liver?

A

bilirubin

bile acids

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11
Q

what are the main components assessed when looking at the liver?

A

enzymes and function markers

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12
Q

levels of what substances produced in the liver are tested on a biochemistry panel?

A

cholesterol, urea, albumin, coagulation factors

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13
Q

what are liver parameters measured in?

A

level of increase above the reference interval (e.g. 10x)

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14
Q

what enzyme level can be elevated in dogs without cholestasis?

A

ALP is increased by steroids and phenobarbital

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15
Q

what can AST levels be increased by as well as liver damage?

A

muscle damage

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16
Q

what is measured to assess kidney function?

A

urea

creatinine

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17
Q

what is measured to assess protein levels?

A

TP, albumin

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18
Q

what is measured to assess electrolyte levels?

A

Na, K, Cl, Ca, PO4

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19
Q

what is measured to check blood sugar levels?

A

glucose

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20
Q

what is measured to assess lipid levels?

A

triglycerides

cholesterol

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21
Q

what does quality control ensure?

A

results are reliable

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22
Q

what should be involved in ensuring quality control?

A 
set up of machines
maintenance and cleaning of machines
interpretation of results 
checks to ensure values are acceptable (control test)
recording
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23
Q

what does the level of cleaning of biochemistry machines depend on?

A

level of use - if used frequently then may not need to be cleaned every time

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24
Q

what should be checked during the maintenance of biochemistry machines?

A

temperature and pH of reagents and that they are up to date. This ensures reactions happen properly and enzymes have optimum working conditions

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25
Q

why is quality control of biochemistry results so important?

A

decisions may be made based on wrong values - even euthanasia
results may have been wrong for a prolonged period of time without anyone being aware

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26
Q

what are the 2 main variables which affect test results?

A

biological

analytical

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27
Q

what are the 2 key biological factors which can affect test results?

A

inter-individual

intra-individual

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28
Q

what are the 3 analytical factors which can affect test results?

A

pre-analytical (before)
analytical (during)
post-analytical (after)

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29
Q

what variables will affect the result most pre-test?

A

biological and pre-analytical

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30
Q

what are inter-individual variables?

A

inherent differences between groups of animals due to the effects of species, breed, age and sex

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31
Q

what is an example of a common inter-individual variable associated with species?

A

cats have lower PCV than dogs

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32
Q

what are common inter-individual variables associated with age?

A

growing dogs have higher Ca, phosphate, and ALP but lower total protein concentration than adult dogs (this will also differ between breeds)

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33
Q

what are intra-individual variables?

A

transient differences in the same animal due to environment/external factors

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34
Q

what are examples of environmental/external factors that will form intra-individual variables?

A

diet, excitement, reproductive status, drugs, method of sampling

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35
Q

what factors should be minimised?

A

intra-individual

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36
Q

how can intra-individual diet factors be minimised?

A

all patients fasted for 8-12 hours pre biochemistry

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37
Q

what are the main pre-analytical factors that affect biochemistry results?

A 
poor sampling/type of sampling 
haemolysed, lipaemic or icteric plasma
wrong anticoagulant
wrong anti-coagulant/blood ratio
wrong tube
transport of sample
storage
contamination with EDTA
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38
Q

in what order must anticoagulant/blood ratios be respected?

A

clotting
haematology
biochemistry

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39
Q

how should samples be stored/transported?

A

refrigerated not frozen unless serum/plasma are separated

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40
Q

how can contamination with EDTA be avoided?

A

don’t touch tube when transferring sample from the clotting test tube as it will ruin biochemistry tests

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41
Q

what method may give a falsely low reading during enzyme testing?

A

kinetic (enzymatic) assays instead of end point assays

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42
Q

what is the difference between end point and kinetic (enzymatic) assays?

A

kinetic (enzymatic) assays measure activity rather than amount of enzyme at the pre-determined end point

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43
Q

why may kinetic (enzymatic) assays give a false low?

A

if there is high amount of enzyme which leads to substrate depletion and so reduced enzyme activity

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44
Q

how can false lows in kinetic (enzymatic) assays be retested?

A

diluting solution

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45
Q

what are reference intervals?

A

most prevalent value in the population

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46
Q

what percentage of the population fall inside reference intervals?

A

95% of population

1 in 20 will have readings outside RI and be healthy

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47
Q

as as well between patients where will reference intervals vary?

A

between biochemistry machines

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48
Q

describe the pattern of changes in liver enzymes due to acute hepatic injury

A

ALT is most sensitive and shows initial increase, will also be the last to decrease
AST is less sensitive as it is found within the mitochondria of hepatocytes

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49
Q

describe the pattern of changes in liver enzymes due to cholestasis

A

cholestatic enzymes will increase to deal with increase bile presence (blockage preventing drainage)
if issue is prolonged then bile will begin to cause damage leading to increase in damage enzymes

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50
Q

what is an increase in bilirubin known as?

A

hyperbilirubinaemia

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51
Q

what does hyperbilirubinaemia lead to?

A

jaundice

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52
Q

what is bilirubin?

A

breakdown product of heamoglobin in RBC

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53
Q

where is bilirubin processed and excreted?

A

processed in liver and excreted in the faeces through bile

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54
Q

what are the 3 causes of hyperbilirubinaemia?

A

pre-hepatic: increased RBC breakdown
hepatic: decreased bilirubin processing (e.g. liver disease)
post-hepatic: decreased bilirubin excretion (e.g. bile duct obstruction)

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55
Q

where are bile acids produced and excreted?

A

in the liver and excreted into small intestine via bile duct

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56
Q

where are bile acids reabsorbed?

A

in distal small intestine

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57
Q

what is reabsorption of bile acids known as?

A

enterohepatic recycling

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58
Q

what are the 3 main reasons for bile acid elevation?

A

decreased hepatic function
decreased bile flow (cholestasis)
portosystemic shunt

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59
Q

what is a portosystemic shunt?

A

flow of blood through portal vein to liver enters the systemic circulation instead

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60
Q

what does a jaundiced sample suggest about bile acids?

A

they will be high

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61
Q

describe the bile acid stimulation test

A

fast overnight, take fasted sample into a plain tube, feed, take a sample into a plain tube 2 hours after feeding

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62
Q

what must be checked before sample collection?

A

tube type, volume of blood/plasma required for each test

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63
Q

why does the volume of blood not equal the volume of plasma?

A

PCV makes up around 45% of blood volume in dogs which is not usable for biochemistry

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64
Q

how much blood should you aim to collect when sampling?

A

3x the amount of plasma volume required - allows for PCV and less complaint patient

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65
Q

how can haemolysis and platelet clumping be avoided in sampling?

A

clean stick

avoid vacuum and collapse of veins by not drawing back on syringe too quickly

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66
Q

what tubes can be used for biochemistry tests?

A

plain tube/serum
heparin tube
Oxf tube

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67
Q

what should Oxf tubes be used for?

A

measurement of glucose when exact values needed as Oxf stops RBC continuing to use glucose

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68
Q

what is happening to glucose in plain and heparin tubes?

A

cells are still living and so glucose is still being consumed

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69
Q

what does lipaemia lead to visibly?

A

turbid/milky serum/plasma

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70
Q

what is lipaemia due to?

A

presence of lipids

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71
Q

what effect will lipaemia have on values?

A

will increase and decrease some values in plasma/serum due to extra lipid fractions and turbidity caused by lipids

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72
Q

what will ideal biochemistry systems do with regards to lipaemia?

A

warn of presence and grade it

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73
Q

does lipaemia affect hematology?

A

yes

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74
Q

what does lipaemia affect that influences results?

A

light transmission which will affect spectrophotometric assays)
can result in apparent dilution of normal substances in aqueous component leading to a false low (e.g. electrolytes)

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75
Q

what are the physiological reasons for lipaemia?

A

post-prandial (need fasted samples!!)

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76
Q

what are the pathological reasons for lipaemia?

A

endocrinopathies e.g. diabetes, hypothyroidism, hyperadrenocortisolism

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77
Q

will pathological lipaemia dissappear when fasted?

A

no

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78
Q

what does haemolysis cause?

A

red coloured serum/plasma due to free haemoglobin/myoglobin

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79
Q

what may cause haemolysis?

A

in vitro - pathological

in vivo - improper handling/sampling most common

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80
Q

what does haemolysis increase in biochemistry?

A

plasma/serum values of some compounds/enzymes due to increased concentration of RBC

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81
Q

what does haemolysis decrease in biochemistry?

A

plasma/serum values of compounds due to decreased concentration of RBC

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82
Q

what does haemolysis interfere with?

A

determinations of colour interference (spectrophotometric assays) or chemical interactions
haematology

83
Q

what is icterus?

A

yellow serum/plasma

84
Q

what effect does jaundice have on biochemistry?

A

depends on the biochemistry being done

85
Q

what is icterus/jaundice due to?

A

increased bilirubin - pathological

86
Q

does jaundice interfere with haematology?

A

no

87
Q

does icteric plasma/serum interfere with assays?

A

yes - should be rerun with a hopefully better sample

88
Q

what are the main 5 types of in-house biochemistry analysers?

A

glucometres
‘dry’ biochemistry analysers (reagents are dry)
‘wet’ biochemistry analysers (reagents are wet)
electrolyte analyser
blood gas analyser

89
Q

describe the basic principle of spectrophotometry

A

light source is passed through the sample and based on how much light of a particular wavelength is absorbed we can calculate concentration of the compound of interest

90
Q

what law is spectrophotometry based on?

A

beers law

absorbance = constant + light pathway distance through cavette - concentration of substance of interest

91
Q

what factors can interfere with spectrophotometry?

A

haemolysis and lipaemia can affect how much light is absorbed and can affect accuracy of results

92
Q

how can haemolysis and lipaemia be prevented?

A

fasting and careful collection/handling

93
Q

describe the best way to prevent haemolysis at all steps

A

careful sampling technique
avoid delays to sample processing
separate plasma/serum from sample as early as possible
refrigerate separated sample

94
Q

what parameters are used to measure the function of the pancreas?

A

PLI and TLI

95
Q

what does PLI stand for?

A

pancreatic lipase immunoreactivity

96
Q

what does TLI stand for?

A

tripsenogen like immunoreactivity

97
Q

what does elevated PLI indicate?

A

pancreatic injury

98
Q

what does TLI indicate?

A

pancreatic functional mass - how much exocrine function remains (EPI)

99
Q

what is EPI?

A

exocrine pancreatic insufficiency

100
Q

why are the bile duct and pancreas so often linked in illness?

A

bile and pancreatic juice drain through a common duct. If there is cholestasis this can lead to secondary pancreatitis and vice versa

101
Q

what 3 parameters are measured to show kidney function?

A

urea
creatinine
SDMA

102
Q

what does increased urea in a biochemistry screen indicate?

A

dehydration, renal disease, urinary obstruction/rupture, heart disease

103
Q

what does decreased urea in a biochemistry screen indicate?

A

liver failure as urea is made in the liver

104
Q

what does increased creatinine in a biochemistry screen indicate?

A

as with urea: dehydration, renal disease, urinary obstruction/rupture, heart disease in medium and large dogs

105
Q

what does decreased creatinine in a biochemistry screen indicate?

A

muscle wastage (very small dogs and cats)

106
Q

is SDMA affected by muscle mass?

A

no, unlike creatinine

107
Q

what must urea, creatinine and SDMA be assessed in conjunction with?

A

urine analysis

108
Q

what do urea, creatinine and SDMA show?

A

glomerular filtration rate

109
Q

how are urea, creatinine and SDMA linked?

A

tend to increase and decrease together

110
Q

define azotemia

A

increased serum urea and/or creatinine concentration

111
Q

where is azotemia detected?

A

in the blood

112
Q

what are the 3 types of azotemia?

A

pre-renal
renal
post-renal

113
Q

what can cause pre-renal azotemia?

A

decreased renal perfusion (hypovolaemia, dehydration)

114
Q

what can cause renal azotemia?

A

disease - only seen when 60-70% of nephrons are damaged

115
Q

what causes post-renal azotemia?

A

urinary tract obstruction or rupture

116
Q

define uremia

A

specific clinical syndrome (vomiting, anorexia and ulcreation)

117
Q

when does uremia develop?

A

when high levels of toxic substances (e.g. urea, creatinine)are present in the blood (azotemia) which causes reduced renal excretion

118
Q

what is CK enzyme linked with?

A

muscle damage

119
Q

what would increased CK indicate?

A

skeletal muscle damage or haemolysis

120
Q

what other parameters may be increased alongside CK?

A

ALT and AST as they are also found within muscle

121
Q

what is the totel proteins measurement formed of?

A

albumin + globulin

122
Q

where is albumin produced?

A

liver

123
Q

how can total proteins be measured?

A

refractometry

biochemistry analyser

124
Q

what are the issues with refractometry when calculating total proteins?

A

may also measure glucose, cholesterol, urea etc, and is affected by lipaemia

125
Q

what information about total protein will be given by biochemistry analyser?

A

albumin value and total protein

126
Q

how can globulin be calculated from biochemistry analyser results?

A

total protein - albumin = globulin

127
Q

what is the role in the blood of proteins?

A

maintaining oncotic pressure as well as individual protein functions

128
Q

how may age affect total proteins?

A

increased in neonates after colostrum ingestion, decreases as maternal antibodies drop

129
Q

what is the most common protein in the blood?

A

albumin

130
Q

what can cause increased albumin in the blood?

A

dehydration

131
Q

what can cause reduced albumin in the blood?

A 
mild inflammation
liver disease
kidney disease
GI disease 
haemorrhage
132
Q

what is SPE?

A

serum protein electrophoresis

133
Q

what is SPE used for?

A

evaluating the different groups of protein in serum

134
Q

what do protein fractions migrate according to, during SPE?

A

size and charge

135
Q

what can cause high glucose?

A

diabetes
stress
steroid therapy

136
Q

what can cause low glucose?

A 
wrong blood tube/time delay
insulinoma
insulin overdose
some tumors (very metabolically active)
137
Q

what does fructosamine indicate?

A

glucose level for past 2-3 weeks

138
Q

what is fructosamine used to diagnose?

A

diabetes

139
Q

what are the common liver parameters measured during biochemistry analysis?

A

ALT, GLDH, ALP, GGT, bilirubin, bile acids

140
Q

what are the common kidney parameters measured during biochemistry analysis?

A

urea, creatinine, SDMA and urine analysis

141
Q

what are the common pancreas parameters measured during biochemistry analysis?

A

PLI, TLI

142
Q

what are the common muscle parameters measured during biochemistry analysis?

A

CK, AST

143
Q

what are the common protein parameters measured during biochemistry analysis?

A

TP, albumin, APPs (acute phase proteins)

144
Q

what are the common electrolyte parameters measured during biochemistry analysis?

A

Na, K, Cl, Ca, PO4

145
Q

what are the common glucose parameters measured during biochemistry analysis?

A

fructosamine

146
Q

what are the common lipid parameters measured during biochemistry analysis?

A

triglycerides and cholesterol

147
Q

what are the common hormone parameters measured during biochemistry analysis?

A

T4, cortisol, TSH, ACTH, insulin

148
Q

what are the common haematology parameters measured during biochemistry analysis?

A

RBC, WBC, Plt, blood smear

149
Q

what are the common haemostasis parameters measured during biochemistry analysis?

A

PT, aPTT, TCT, BMBT

150
Q

what can cause increased total proteins?

A

false high with lipaemia
dehydration
inflammation
neoplasia

151
Q

what can cause decreased total proteins?

A 
haemorrhage
GI disease
very young animals
renal disease
hepatic disease
152
Q

which conditions have the largest effect on albumin?

A

renal disease

hepatic disease

153
Q

what is haemostasis?

A

the ability to stop bleeding

154
Q

what does defective haemostasis lead to?

A

leads to haemorrhage, generally known as coagulopathies

155
Q

what does excessive haemostasis lead to?

A

leads to thrombotic events

156
Q

what anticoagulant is used for clotting tests?

A

EDTA

157
Q

what are the most important considerations when performing clotting tests?

A

respect amount of blood required
perform clean venapuncture
separate plasma ASAP
freeze if sending away

158
Q

what are the 3 stages of haemostasis?

A

primary
secondary
tertiary

159
Q

when does primary haemostasis occur?

A

first stage, occurs when the needle punctures the vein and lasts seconds to minutes

160
Q

what is primary haemostasis associated with?

A

platelet numbers, function and vascular function leading to the formation of the platelet plug

161
Q

what does primary haemostasis require?

A

enough platelets
platelets to function properly
von Wilburn factor

162
Q

what happens during secondary haemostasis?

A

stabilisation of clot in minutes to hours

163
Q

what is secondary haemostasis associated with?

A

formation of fibrin mesh which stabilises platelet plug

164
Q

what happens during tertiary haemostasis?

A

return to normal vascular flow

165
Q

what is tertiary haemostasis associated with?

A

breakdown of the clot

166
Q

what can be used to measure tertiary haemostasis?

A

D-Diamers

167
Q

what are the requirements for normal primary haemostasis?

A

vessel wall
platelets in correct number and functionin gproperly
vWF

168
Q

what are the tests available for primary haemostasis?

A

BMBT
platelet count
platelet function testing
histopathology

169
Q

what are the common disorders of primary haemostasis?

A

IMTP - immune mediated thrombocytopenia
vW disease
angiostrongylus

170
Q

what are the requirements for normal secondary haemostasis?

A

coagulation factors
calcium
vitamin K
fibrin

171
Q

what are the tests available for secondary haemostasis?

A

Activated Clotting Time
Prothrombin Time
Activated Partial Thromboplastin Time
Thrombin Clotting Time

172
Q

what are the common disorders of secondary haemostasis?

A

congenital factor deficiencies
warfarin toxicity
severe liver disease
angiostrongylus

173
Q

what are the requirements for normal tertiary haemostasis?

A

tissue plasminogen activator (tPA)

plasmin

174
Q

what are the tests available for tertiary haemostasis?

A

FDPs

D-Diamers

175
Q

what are the common disorders of tertiary haemostasis?

A

DIC

176
Q

What is BMBT?

A

buccal mucosal bleeding time

177
Q

how is BMBT tested?

A

under sedation - standardised cut made on patients gum. Blood collected with filter paper, taking care not to disturb the clot, and is absorbed until it stops

178
Q

what are the normal BMBT for cats and dogs?

A

normal dog <3.3 mins
GA dog <4 mins
GA cat <3.3 mins

179
Q

what causes prolonged BMBT?

A

moderate to severe thrombocytopenia
thrombopathia - inherited or acquired
vWM disease

180
Q

what screening can BMBT be used for?

A

pre op of breeds at risk of vWD e.g. doberman

181
Q

how quickly will hemorrhage start after trauma in a patient with primary haemostasis disorders?

A

immediately

182
Q

what type of bleeding is typically seen in a patient with primary haemostasis disorders?

A

capillary bleeding: skin, mucosa, epistaxis, GI and urinary tract.
petechail haemorrhage
ecchymosis (bruising)

183
Q

how quickly will hemorrhage start after trauma in a patient with secondary haemostasis disorders?

A

usually a small delay

184
Q

what type of bleeding is typically seen in a patient with secondary haemostasis disorders?

A

internal bleeding: joints, muscles, body cavity (pleural, mediasteinum)
haematomas
haemoartroses

185
Q

what bleeding is typically seen in a patient with both primary and secondary haemostasis disorders?

A 
GI bleeding
meleana
haematoemesis
haematochezia
epistaxis
haematauria
186
Q

what is thrombocytopenia?

A

low platelets

187
Q

how is activated clotting time tested?

A

whole blood is collected and allowed to clot in commercially available tubes

188
Q

what are the normal activated clotting time values for dogs?

A

<90 sec

189
Q

what are the normal activated clotting time values for cats?

A

<60 sec

190
Q

when is activated clotting time prolonged?

A

if there is a marked deficiency in one/several clotting factors
affected by platelet number and function

191
Q

what does the clotting cascade result in?

A

fibrin (clot)

192
Q

what are the 2 branches of the clotting cascade

A

extrinsic

intrinsic

193
Q

what can be used to measure extrinsic pathway of the clotting cascade?

A

prothrombolin time (PT)

194
Q

what can be used to measure intrinsic pathway of the clotting cascade?

A

activated partial thromboplastin time (APTT)

195
Q

what are PT and APTT tests checking for?

A

presence of clotting factors and function

196
Q

what causes prolonged PT and APTT tests?

A

Increase in clotting times due to reduction in clotting factor (if individual factor is reduced to <20-30% of normal concentration)

197
Q

where are most clotting factor and cofactors made?

A

in the liver

198
Q

where do most clotting factors and cofactors circulate?

A

in the blood

199
Q

what can be used to test for fibrinolysis?

A

FDPs

D Diamers

200
Q

why to FDPs elevate?

A

increase on clot formation and lysis as well as inflammation

201
Q

why do D Diamers elevate?

A

increases with clot formation and lysis

202
Q

what are D Diamers?

A

degradation products of cross linked fibrin (stable clot)

203
Q

when may elevated D Diamers be seen?

A

post surgery/trauma
DIC secondary to inflammation
neoplasia
thromboembolytic disease

X Pre Clinical Science (17 decks)

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