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Analytical methods > Principles of analysis > Flashcards

Principles of analysis Flashcards

1
Q

What are the 3 phases of testing?

A

pre analytical
analytical
post analytical

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2
Q

Match the following

A. Pre analytical
B. Analytical
C. Post analytical

  1. reference ranges, diagnostic sensitivity and specificity
  2. Methodology, calibraiton, assessment of assay performance, quality control
  3. sample variables and patient variables
A

C1, B2, A3

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3
Q

What is a definitive method?

A

Method of exceptional scientific summary suitable for certification of reference material

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4
Q

What is a reference method?

A

Method demonstrating small inaccuracies against definitive method.

Method to test the performance of the definitive method

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5
Q

What is a routine method?

A

Method deemed sufficiently accurate for routine use against reference method and standard reference materials.

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6
Q

Match the following methodology terms?

A. Reference method
B. Definitive method
C. Routine method

  1. Method of exceptional scientific summary suitable for certification of reference material
  2. Method deemed sufficiently accurate for routine use against reference method and standard reference materials.
  3. Method demonstrating small inaccuracies against definitive method.
A

C2, B1, A3

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7
Q

Use reference, definitive, routine method with an example of cholesterol

A

Definitive method: isotope dilution/gas chromatography/mass spectrometry

Reference method: abell- Kendall Method

  • hydrolysis of cholesterol esters with alcoholic KOH
  • extraction of total cholesterol with hexane for 15 mins -> dried in vacuum
  • treated with acetic acid/acetic anhydride/sulphuric acid 30min then abs at 620nm

Routine method: enzymatic e.g. Beckman

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8
Q

What is a primary standard?

A

Substance of known chemical composition and high purity that can be accurately quantified and used for assigning values to materials and calibrating apparatus

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9
Q

What is standard reference material (SRM)?

A

Reference material issued by an institute whose values are certified by a reference method which establishes traceability

aka. Material produced which has been established as useful by reference method and established traceability

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10
Q

What is a secondary standard?

A

A commercially produced standard for routine use calibrated against a primary standard or reference material

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11
Q

What can calibrators give us clinicians?

A

Allows us to produce a standard curve when we are measuring something being emitted. This means we can confidently quantify the amount of a certain substance.

Allows the relationship of analyte conc and signal to be examined

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12
Q

Requirements of calibrators.

A

Prepared from pure substance
Stable and homogenous material
Matrix similar to assay matrix e.g. serum
If possible should be obtained commercially to min error

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13
Q

Match the following, calibrator values:

A. Stated value
B. Assigned value
C. Certified value
D. Standard reference method value

  1. derived using a reference method
  2. No certification
  3. Given arbitrarily or derived using non-reference method
  4. certification of value by particular institute or body
A

D1, B3, C4, A2

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14
Q

Define traceability

A

An unbroken chain of comparisons of measurements leading to a reference value

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15
Q

Importance of traceability

A

Ensures reasonable agreement between routine and reference method

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16
Q

What makes a good method?

A

Analytical accuracy

Analytical precision

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17
Q

Match
A. Analytical accuracy
B. Analytical precision

  1. Measure of agreement between a measured quantity and true value.
  2. Measure of agreement between replicates.
A

A1, B2

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18
Q

How is precisiona and accuracy implicated in screening?

A

Important to avoid FN and FP

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19
Q

Accuracy is a combination of what and involves what?

A

combination of trueness and precision

involves systemic and random error

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20
Q

What is a random error?

A

Random deviation from expected value due to inaccurate pipetting, sample contamination

21
Q

Systemic error

A

Shift in expected values indicating bias

22
Q

What does trueness measure?

A

systemic error or bias

23
Q

How can degree of precision can be measured?

A

by quantifying the overall effect of all random errors

24
Q

Mehtods of testing trueness?

A

Recovery experiment
Comparison to fresh EQA material
Correlation with a current/accepted method using patient samples

25
Q

Value for precision?

A

Coefficient of variation = SD/M x100%

  • Allows comparison between results from different analyte in a way stdev cant
  • Ideal CV <5%
26
Q

What is an interference?

A

Specific component that can be identified as causing an effect

27
Q

Difference between serum and plasma sample collection.

A

Serum is a clotted blood sample and is therefore devoid of clotting factor (no anticogulant). Plasma anticoagulated before centrifuge

28
Q

Why is blood an appropriate sample for testing?

A

Convient, taken randomly

Detect; leakage from damage tissue, hormones, waste, metabolites not properly controlled, nutrient levels

29
Q

Urine as a sample?

A

Natural waste product of body
Measures: electrolyrs, nitrogenous compounds, phosphates, drug metabolims

Levels of hydration; ratio e.g. to creatinine
Dehydrated indicated by low creatinine

30
Q

Cerebral spinal fluid as sample?

A

Difficult to obtain, but very good for specific conditions
- secretory product of ventricels, choroid plexus

Levels of typical components are similar to those found in blood or plasma

31
Q

What are some blood preservatives and what do they do?

A

Required to prevent metabolism of certain analytes in vivo- can interfere with assays

  • potassium EDTA (anticoagulant)
  • lithium heparin (blocks clotting)
  • sodium citrate
  • fluoride oxalate
32
Q

What is haemolysis?

A

Disruption of the membrane of red blood cells resulting in release of cellular contents - most common intereference

33
Q

What percentage of people fall within +/-1std off the mean?

A

68%

34
Q

What percentage of people fall within +/-2std off the mean?

A

95% this is within the reference range

35
Q

Match the following calculations of reference ranges.

A. Parametric method
B. Non-parametric-method
C. Target driven

  1. Makes no assumption about type of distribution
  2. Reference range cant be derived from healthy pop
  3. Assumes gaussian distribution
A

A3, B1, C2

36
Q

What is excluded in a reference range production?

A

People with Disease, Risk factors, Intake of pharmacological agents, physiological states

37
Q

What is the total CV calculation?

A

CV(T)=sqrt CV^2(ana)+CV^2(biological)

38
Q

Define clinical sensitivity

A

Ability of a test to correctly identify those who have the disease - highly sensitive = few false negatives

39
Q

Define clinical specificity

A

Ability of a test to correctly identify those who do not have a disease
- highly specific = few false positives

40
Q

What is predictive value?

A

measure of the ability of test to correctly assign individual to either disease or non-diseased group

41
Q

What is diagnostic efficiency?

A

proportion of true results

42
Q

Match the following to what describes the the best.

A. Clinical specificity
B. Clinical sensitivity
C. Diagnostic efficiency
D. Predictive value

  1. Proportion of true results
  2. Ability of test to correctly ID those without disease
  3. Ability of test to correctly ID people with disease
  4. Ability of test to correcltly assign either disease or non
A

A2
B3
C1
D4

43
Q

With regards to the 2x2 table how is sensitivity, specificity, positive/negative predictive value and diagnostic efficiency calculated

A 
Sensitivity = TP/(TP+FN)
Specifity = TN/(TN+FP)

PPV= TP/(TP+FP)
PPN=TN/(TN+FN)

44
Q

How is analytical quality monitored as a whole?

A

In real time using an internal quality control (IQC)

Retrospectively through the use of external quality assurance (EQA)

45
Q

Explain IQC.

A

IQC monitors the performance of an assay. Will the same assay produce the same result multiple times? The expected value should be known and if its not obtained the assay should be reconsidered before releasing results to a patient

46
Q

Explain EQC

A

Allows for retrospective analysis of performance of an assay between other users. Are the results the same in other labs using same analyser or assay?

47
Q

What are some basic parameters involved in choosing IQC material?

A

Matrix
- close matrix match. Human were possible

Third party
- min one QC should not be made by assay manufacturer. To minimise bias.

Levels available
- min 2- one normal(physiological) and high (pathological)

Range of analytes
- QC multiple assays

Cost - cost effectiveness without compromising quality

Stability - repeatability

48
Q

How often is IQC needed?

A

Depend on frequency of analysis

  • Batch analysis: required at beginning and end
  • continuous flow requires throughout

Accuracy of results between acceptable IQC results

Depends on analyte

Analytical methods (15 decks)

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