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Microbiolgical Data
Tim Sandle
Microbiology information
resource:
http://www.pharmamicroresources.com/
Introduction
Distribution of microbiological data
Use of trend charts
Calculation of warning and action
levels
Introduction
Examples from environmental
monitoring and water testing
Broad and illustrative overview
Written paper with more detail
Distribution of microbiological data
Why study distribution?
• Impact on sampling
• Impact on trending
• Impact upon calculation of warning and
action levels
Distribution
Most statistical methods are based
on normal distribution, and yet….
Most microbiological data does NOT
follow normal distribution
Distribution
Micro-organisms, such as those in a
typical, free-flowing water system,
follow Poisson distribution
For example…
Distribution
S1 S2 S3
S4 S5
Where S = sample
= micro-organism
Distribution
And microbial counts tend to be
skewed (or positive or negative
exponential distribution)
For example, a Water-for-Injection
system…
Distribution
350
Number of samples
300
250
200
150
100
50
0
1 2 3 4 5 6 7
Count (cfu / 100 ml)
Distribution
So, what can we do about it?
CI for Mean
0 2 4 6 8
Count
Distribution
Whereas if the square root is taken:
Distribution
We move closer to normal
distribution:
CI for Mean
Count
Distribution
Logarithms work in a similar way for
higher counts
Remember to add ‘+1’ to zero counts
(and therefore, +1 to all counts)
Trend Analysis
There is no right or wrong approach
There are competing systems
This presentation focuses on two
approaches, both described as
‘control charts’:
• The cumulative sum chart
• The Shewhart chart
Trend Analysis
Control charts form part of the
quality system
They can be used to show:
• Excessive variations in the data
• How variations change with time
• Variations that are ‘normally’ expected
• Variations that are unexpected, i.e.
something unusual has happened
Trend Analysis
Control charts need:
• A target value, e.g. last year’s average
• Monitoring limits:
Upper limit
Lower limit
Control line / mean
So the data can be monitored over time and
in relation to these limits
Trend Analysis
Of these,
• The warning limit is calculated to represent a
2.5% chance
• The action level is calculated to represent a
0.1% chance
• So, if set properly, most data should remain
below these limits
• These assumptions are based on NORMAL
DISTRIBUTION
• Various formula can be used to set these or
validated software
Trend Analysis
Cumulative sum chart (cusum)
• Suitable for large quantities of low count
data. It is very sensitive to small shifts
• Shows shifts in the process mean
Shewhart chart
• Suitable for higher count data. It shows
large changes more quickly.
Trend Analysis
Cusums
• Harder to interpret
• Displays the cumulative sum of a rolling
average of three values and plots these
in comparison with the target value
• The direction and steepness of the slope
are important
• Significant changes are called ‘steps’
• V-masks can be used as a prediction to
the future direction
Trend Analysis
For example, a Grade B cleanroom
Contact (RODAC) plates are
examined
A target of 0.2 cfu has been used,
based on data from the previous
year
Trend Analysis
Trend Analysis
Shewhart charts
• Powerful for distinguishing between
special causes and common causes
• Common causes are inherent to the
process and are long-term
• Special causes are where something has
changed and maybe of a long or short
term
Trend Analysis
Examples of special causes:
• a) A certain process
• b) A certain outlet
• c) A certain method of sanitisation, etc.
• d) Sampling technique
• e) Equipment malfunction e.g. pumps, UV
lamps
• f) Cross contamination in laboratory
• g) Engineering work
• h) Sanitisation frequencies
Trend Analysis
For example, a Grade C cleanroom
• Active air-samples are examined
• A target of 1.5, based on historical data
Trend Analysis
Trend Analysis
The previous charts were prepared
using a statistical software package
However, MS Excel can also be used
The next example is of a WFI system
Notice the data has been converted
by taking the square root of each
value
Trend Analysis
3.5
Sq root of mean count /
3
2.5
2
week
1.5
1
0.5
0
13
17
21
25
29
33
37
41
45
49
53
57
61
1
5
9
-0.5
-1
Number of weeks
Trend Analysis
Alternatives:
• Individual Value / Moving Range charts
• Exponentially Weighted Moving Average
charts (EWMA)
• These are useful where counts are NOT
expected, e.g. Grade A environments
• They look at the frequency of intervals
between counts
Trend Analysis
Summary