welcome

Preparation of control sera

 Types and Sources of Control Materials.
 Serum collection and screening for
infectious diseases.
 Preparation of different control levels.
 Analysis of Control Materials.
 Calculation.
 Summary comments

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Quality Control for
Quantitative Tests
Preparation of
control sera
Majid Mohammed Ali
M.Sc Clinical Chemistry
 Sources for Control Materials
Commercial product.
Diagnostic samples (qualitative QC).
“Homemade” or “In-house”.
Obtained from:
Another laboratory .
EQA provider.

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 Quantitative QC Materials
• Calibrator:
• a solution which has a known amount of
analyte weighed in or has a value
determined by repetitive testing using a
reference or definitive test method
• Control:
• material or preparation used to monitor
the stability of the test system within
predetermined limits

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 Control solutions- control
materials
The International Federation of
Clinical Chemistry defines a
control solution or control material
as a "specimen or solution which
is analyzed solely for quality
control purposes, not for
calibration" .
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 Control solutions- control
materials
We use the term control material
or control product to refer to a
control solution that is available,
usually commercially, in liquid,
frozen, or lyophylized form,
packaged in small bottles suitable
for use on a daily basis.
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 Control Material

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 Types
Types of
of Control
Control Materials
Materials
 Assayed
mean calculated by the manufacturer
must verify in the laboratory
 Unassayed
less expensive
must perform data analysis
 “Homemade” or “In-house”
pooled sera collected in the laboratory
characterized
preserved in small quantities for daily use

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 Assayed versus Unassayed
controls
 Control products are available as assayed
or Unassayed materials.
 Assayed control materials generally
come with an assay sheet of expected
values for analytes assayed by various
methods and instrument systems.
 These assay sheets usually list, for each
constituent present, expected mean
values as well as expected ranges.
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 “Homemade” or “In-house”
pooled sera
pooled sera collected from:

pooled sera collected in the

laboratory.
pooled sera collected in the blood
bank.
pooled sera collected from animals.

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 In-house pooled sera
preparation
Collection.
Screening for infectious
diseases (HIV 0,1,2, HBsAg,
HCV, Syphilis).
Freezing to -70 °C( 24 Hr).

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 Analyte levels

Constituent levels of quality

control materials should be
chosen at medical decision
concentrations and/or at critical
method performance limits such
as upper and lower linearity
limits.
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 Analyte levels
Two or three different concentrations
are often needed for each analyte.
 Choosing control materials at critical
concentrations will allow the analyst
to estimate the random error at critical
levels of the method .

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 levels of quality control
materials
Transfer the cooled container to room
temperature.
Put it upside down on clean dry beaker.
Separate to three plasma fraction from
the freeze pooled sera.
Separated pooled sera fraction stabilized
with ethanediol.

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 Analysis
Analysis of
of Control
Control Materials
Materials
 Need data set of at least 20 reading to each
parameter.
 Make sure any procedural variation is
represented:
different operators
different times of day
Different instruments.
 Determine the degree of variability in the
data
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to establish acceptable
Majid Mohammed Ali
range. 16
 Calculation
oCalculate mean.
o Standard deviation.
o Coefficient of variation.
oDetermine target ranges.

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 Calculation
Calculation of
of Mean
Mean

(X ) = X +X +X
1 2 3
... + X n
n
X = Mean
X1 = First result
X2 = Second result
Xn = Last result in
series
n – Total number of
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results
 Calculation
Calculation of
of Mean
Mean to
to glucose
glucose results
results

1. 192 mg/dL 7. 200 mg/dL

2. 194 mg/dL 8. 200 mg/dL
3. 196 mg/dL 9. 202 mg/dL
4. 196 mg/dL 10. 255 mg/dL
5. 160 mg/dL 11. 204 mg/dL
6. 196 mg/dL 12. 208 mg/dL
13. 212 mg/dL

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 Calculation
Calculation of
of Mean
Mean to
to glucose
glucose results
results
1) 192 mg/dL
2) 194 mg/dL
3) 196 mg/dL
4) 196 mg/dL
5) 196 mg/dL • Mean = the calculated
6) 200 mg/dL average of the values
7) 200 mg/dL
8) 202 mg/dL • The sum of the values (X1
9) 204 mg/dL + X2 + X3 … X11) divided
10)208 mg/dL
by the number (n) of
11)212 mg/dL
Sum = 2,200 mg/dL observations
• The mean of these 11
observations is (2200 ÷
11) = 200 mg/dL

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 Calculation
Calculation of
of Variance
Variance
Variance is a measure of variability
about the mean.
It is calculated as the average squared
deviation from the mean.
the sum of the deviations from the
mean, squared, divided by the
number of observations (corrected for
degrees of freedom)

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 Calculation
Calculation of
of Variance
Variance (S2)
(S2)

∑ (X − X )
S = N −1
2 1

= mg /dl
2
2
2

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 Calculation
Calculation of
of Standard
Standard Deviation
Deviation
 The standard deviation (SD) is the square root of the
variance
it is the square root of the average squared deviation
from the mean
 SD is commonly used (rather than the variance) since it
has the same units as the mean and the original
observations
 SD is the principle calculation used in the laboratory to
measure dispersion of a group of values around a
mean.

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 Calculation
Calculation of
of Standard
Standard Deviation
Deviation

(x − x )
S= = mg/dl
2
1

N −1

variance
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 Standard
Standard Deviation
Deviation and
and Probability
Probability
• For a set of data with a
normal distribution, a value X
will fall within a range of:

Frequency
– +/- 1 SD 68.2% of the time
– +/- 2 SD 95.5% of the time 68.2%
– +/- 3 SD 99.7% of the time

95.5%
99.7%
-3s- 2s -1s Mean +1s +2s
+3s

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 Standard
Standard Deviation
Deviation and
and Probability
Probability

 In general, laboratories use the +/- 2 SD

criteria for the limits of the acceptable range
for a test
 When the QC measurement falls within that
range, there is 95.5% confidence that the
measurement is correct
 Only 4.5% of the time will a value fall
outside of that range due to chance; more
likely it will be due to error
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 Calculation
Calculation of
of
Coefficient
Coefficient of
of Variation
Variation

• The coefficient of
variation (CV) is the
standard deviation SD
(SD) expressed as a CV = x 100
percentage of the mean
mean
• Ideally should be
less than 5%

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 Range
Range

o Range refers to the difference or spread

between the highest and lowest
observations.
o It is the simplest measure of dispersion.
o It makes no assumption about the shape
of the distribution or the central tendency
of the data.
o Calculated from the mean ± 2sd.
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 Summary comments

• The selection of appropriate control

materials requires consideration of
many factors and should be part of
the planning process for the selection
and implementation of SQC.
• Compromises may have to be made
in order to limit the number of control
materials used.

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 Summary comments

• There is no right or wrong way to

choose control materials for a
given method, just as there is no
perfect control which behaves
exactly the same as a fresh
human sample.
• Control materials being used
should be reassessed yearly.
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 Summary comments

• Selection of appropriate control

materials is a balancing act in which
cost, stability, ease of use,
performance due to matrix effects,
and constituent levels must be
considered and weighted for each
laboratory’s own application and use.

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 Questions ???

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