QUALITY CONTROL

Amal George
SBS MGU
george.oa.jr@gmail.com
 DEFINITIONS
Quality Control - QC refers to the measures that must be included
during each assay run to verify that the test is working properly.

Quality Assurance - QA is defined as the overall program that ensures

that the final results reported by the laboratory are correct.
“The aim of quality control is simply to ensure that the results
generated by the test are correct. However, quality assurance is
concerned with much more: that the right test is carried out on the
right specimen, and that the right result and right interpretation is
delivered to the right person at the right time”
IMPORTANT TERMINOLOGIES
Quality
The Degree to which a set of inherent characters fulfils the
requirements.
Precision
Precision is the reproducibility of an analytical method

Accuracy
Accuracy defines how close the measured value is to the actual value.
Sensitivity
Sensitivity of an assay in a measure of how little of the analyte the
method can detect.

Specificity
Specificity of a assay related to how good the assay is at discriminating
between the requested analyte and potentially interfering substances.
Standard
This is a substance of constant composition of sufficient purity to be
used for comparison purposes or standardisation
Control
This is a sample, which is chemically and Physically similar to
unknown specimen
A solution , lyophilised preparation or pool of collected human or
animal specimen or artificially derived material intend for use in the QC
process.
Calibrator:
A material or solution of
known concentration (or
activity / intensity) used to
calibrate or adjust a
measurement procedure. It is
also used to calculate the
concentration of an unknown
sample (as standard)
 Quality Control
Variation in results

Biochemical measurements vary for two reasons, namely:

analytical variation
biological variation.
 ANALYTICAL ERRORS
Analytical Measurement
Instrument not calibrated correctly
Specimens mix – up
Incorrect volume of specimen
Interfering substances present
Instrument precision problem

Test reporting
Wrong patient ID
Transcription error
Report not legible
Report delayed
 POST ANALYTICAL ERRORS
Test interpretation
Interfering substance not recognized
Specificity of the test not understood
Precision limitation not recognized
Analytical sensitivity not appropriate
Previous values not available for
comparison
HOW TO CONTROL THESE ERRORS?
PRE ANALYTICAL VARIABLES
It is very difficult to establish effective methods for monitoring and
controlling preanalytical variables because many of the variables are
outside the laboratory areas.
Requires the coordinated effort of many individuals and hospital
departments
Patient Identification
The highest frequency of errors occurs with the use of handwritten labels
and request forms. The use of bar code technology has significantly
reduced ID problems.
Turn around time
Delayed and lost test requisitions, specimens and reports can be major
problems for labs. Recording of the actual times of specimen collection,
receipt in the lab and reporting of results with use of computers will
solve these problems.
Transcription error
A substantial risk of transcription error exists from manual entry of data even
with the double checking of results, computerization will reduce this type of
transcription error.

Patient preparation

Lab tests are affected by many factors, such as,

recent intake of food, alcohol, or drugs
smoking
exercise
stress
sleep
posture during specimen collection
The lab must define the instructions and procedures compliance with these
instructions can be monitored directly efforts should be made to correct non
compliance
GENERAL PRINCIPLES OF CONTROL CHARTS

 Control charts are simple graphical displays in which the

observed values are plotted versus the time when the observations
are made.

 The control limits are calculated from the mean (x) and
standard deviations (s)
Internal Quality Control Program for
Serological Testing
An internal quality control program depend on the use of internal
quality control (IQC) specimens, Shewhart Control Charts, and the use
of statistical methods for interpretation.

Internal Quality Control Specimens

IQC specimens comprises either (1) in-house patient sera (single or

pooled clinical samples), or (2) international serum standards with
values within each clinically significant ranges.
QC Log with Patient Results
 Basic statistics to develop an acceptable control
range
The most fundamental statistics used by the laboratory are the mean [x] and
standard deviation [s].
Calculating a Mean [x]:
The mean (or average) is the laboratory’s best estimate of the analyte’s true
value for a specific level of control.
Calculating a Standard Deviation [s]:
 Standard deviation is a statistic that quantifies how close numerical values
(i.e., QC values) are in relation to each other
 Standard deviation is calculated for control products from the same data
used to calculate the mean.
 It provides the laboratory an estimate of test consistency at specific
concentrations.
 The repeatability of a test may be consistent (low standard deviation, low
imprecision) or inconsistent (high standard deviation, high imprecision).
Mean [x] Standard Deviation [s]
 Creating a Levey-Jennings Chart
• Standard deviation is commonly used for preparing Levey-Jennings (L-J or LJ) charts.
The Levey-Jennings chart is used to graph successive (run-to-run or day-to-day)
quality control values.
• A chart is created for each test and level of control.
• The first step is to calculate decision limits. These limits are ±1s, ±2s and ±3s from
the mean.
• From the mean and standard deviation value calculated in the previous slide we can
now construct the Levy Jennings chart as follows:
The Levey-Jennings chart that was developed can be overlaid onto a bell-shaped curve to illustrate the
overall distribution of quality control values
Cont..
• When an analytical process is within control, approximately 68% of all
QC values fall within ±1 standard deviation (1s).
• Likewise 95.5% of all QC values fall within ±2 standard deviations (2s)
of the mean. About 4.5% of all data will be outside the ±2s limits
when the analytical process is in control. Approximately 99.7% of all
QC values are found to be within ±3 standard deviations (3s) of the
mean.
• As only 0.3%, or 3 out of 1000 points, will fall outside the ±3s limits,
any value outside of ±3s is considered to be associated with a
significant error condition and patient results should not be reported.
 Systematic Errors
Systematic error is evidenced by a change in the mean of the control values. The change in the mean may be gradual
and demonstrated as a trend in control values or it may be abrupt and demonstrated as a shift in control values.

Trends: A trend indicates a gradual loss of reliability in the test system. Trends are usually subtle. Causes of
trending may include:

 Deterioration of the instrument light source

 Gradual accumulation of debris in sample/reagent tubing
 Gradual accumulation of debris on electrode surfaces
 Aging of reagents
 Gradual deterioration of control materials
 Gradual deterioration of incubation chamber
 temperature (enzymes only)
 Gradual deterioration of light filter integrity
 Gradual deterioration of calibration
Shift
Abrupt changes in the control mean are defined as shifts. Shifts in QC data represent a
sudden and dramatic positive or negative change in test system performance. Shifts
may be caused by:
 Sudden failure or change in the light source
 Change in reagent formulation
 Change of reagent lot
 Major instrument maintenance
 Sudden change in incubation temperature (enzymes
only)
 Change in room temperature or humidity
 Failure in the sampling system
 Failure in reagent dispense system
 Inaccurate calibration/recalibration
 Random Errors

 Random error is any deviation away from an expected

result.
 For QC results, any positive or negative deviation away from
the calculated mean is defined as random error.
 There is acceptable (or expected) random error as defined
and quantified by standard deviation.
 There is unacceptable (unexpected) random error that is
any data point outside the expected population of data
(e.g., a data point outside the ±3s limits).
 Westgard Rules

There are six basic rules in the Westgard scheme.

These rules are used individually or in combination to evaluate the quality
of analytical runs.
Most of the quality control rules can be expressed as NL where N
represents the number of control observations to be evaluated and L
represents the statistical limit for evaluating the control observations
Thus 13s represents a control rule that is violated when one control
observation exceeds the ±3s control limits.
Rule 12s Rule 13s
This rule merely warns that random error This rule identifies unacceptable
or systematic error may be present in the
test system random error or possibly the
The relationship between this value and beginning of a large systematic
other control results within the current error. Any QC result outside ±3s
and previous analytical runs must be violates this rule.
examined. If no relationship can be found
and no source of error can be identified,
it must be assumed that a single control
value outside the ±2s limits is an
acceptable random error.
 Rule 22s
This rule identifies systematic error only. The criteria for
violation of this rule are:
• Two consecutive QC results
• Greater than 2s
• On the same side of the mean

There are two applications to this rule: within-run and across runs. The
within-run application affects all control results obtained for the current
analytical run.
• If a normal (Level I) and abnormal (Level II) control are assayed in this
run and both levels of control are greater than 2s on the same side of
the mean, this run violates the within-run application for systematic
error. If however, Level I is -1s and Level II is +2.5s (a violation of the 12s
rule), the Level II result from the previous run must be examined. If
Level II in the previous run was at +2.0s or greater, then the across run
application for systematic error is violated.
Violation of the within-run application indicates that systematic error is
present and that it affects potentially the entire analytical curve.
Violation of the across run application indicates that only a single
portion of the analytical curve is affected by the error.
 Rule R4s

• This rule identifies random error only,

and is applied only within the current
run. If there is at least a 4s difference
between control values within a single
run, the rule is violated for random
error.
• For example, assume both Level I and
Level II have been assayed within the
current run. Level I is +2.8s above the
mean and Level II is -1.3s below the
mean. The total difference between the
two control levels is greater than 4s
(e.g. [+2.8s – (-1.3s)] = 4.1s).
Rule 31s
The criteria which must be met to violate this rule are:
• Three consecutive results
• Greater than 1s
• On the same side of the mean

Rule 41s

The criteria which must be met to violate this rule are:

• Four consecutive results
• Greater than 1s
• On the same side of the mean

There are two applications to the 31s and 41s rule. These are within control material (e.g. all Level I
control results) or across control materials (e.g., Level I, II, and III control results in combination).

Within control material violations indicate systematic bias in a single area of the method curve while
violation of the across control materials application indicates systematic error over a broader
concentration
 Rules 7X 8X 9X 10X12X

These rules are violated when there are: 7 or

8, or 9, or 10, or 12 control results On the
same side of the mean regardless of the
specific standard deviation in which they are
located.

Each of these rules also has two applications:

within control material (e.g., all Level I
control results) or across control materials
(e.g. Level I, II, and III control results in
combination).

Within control material violations indicate

systematic bias in a single area of the method
curve while violation of the across control
materials application indicates systematic
bias over a broader concentration
 Coefficient of Variation [CV]
• The Coefficient of Variation [CV] is the ratio of the standard deviation to the mean and is expressed as a
percentage.
• CV allows the technologist to make easier comparisons of the overall precision.
• Standard deviation typically increases as the concentration of the analyte increases, the CV can be regarded
as a statistical equalizer.
• Technologist/technician who is comparing precision for two different methods and uses only standard
deviation, can be easily misled
• For example, a comparison between hexokinase and glucose oxidase (two methods for assaying glucose) is
required. The standard deviation for the hexokinase method is 4.8 and it is 4.0 for glucose oxidase. If the
comparison only uses standard deviation, it can be incorrectly assumed that the glucose oxidase method is
more precise that the hexokinase method. If, however, a CV is calculated, it might show that both methods
are equally precise. Assume the mean for the hexokinase method is 120 and the glucose oxidase mean is
100. The CV then, for both methods, is 4%. They are equally precise.
 Coefficient of Variation Ratio [CVR]
• Accuracy of test results is paramount in the clinical laboratory, precision is just as
important
• One way a laboratory can determine whether the precision of a specific test is
acceptable is to compare its precision to that of another laboratory performing the same
test on the same instrument using the same reagents (laboratory peer group)
• If the CV for potassium on a particular instrument is 4% and the potassium for all other
laboratories using the same instrument is 4.2%, then the coefficient of variation ratio
[CVR] is 4/4.2 or 0.95.
• Any ratio less than 1.0 indicates that precision is better than the peer group. Any score
greater than 1.0 indicates that imprecision is larger
• Ratios greater than 1.5 indicate a need to investigate the cause of imprecision and any
ratio of 2.0 or greater usually indicates need for troubleshooting and corrective action
 Standard Deviation Index [SDI]
• The standard deviation index [SDI] is a peer-based estimate of reliability. If the peer group mean is
defined as XGroup , the standard deviation is defined as SGroup and the laboratory’s mean is
defined as Xlab
• The target SDI is 0.0 which indicates a perfect comparison with the peer group. The following
guidelines may be used with SDI. A value of:
• 1.25 or less is considered acceptable.
• 1.25 – 1.49 is considered acceptable to marginal performance. Some investigation of
• the test system may be required.
• 1.5 – 1.99 is considered marginal performance and investigation of the test system is
• recommended.
• 2.0 or greater is generally considered to be unacceptable performance and remedial action is
usually required.
Thankyou!