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This method is probably the most familiar one to microbiologists, as the calculation is also used to estimate the initial population of organisms in a sample analyzed by serial dilution techniques.The MPN calculation has different uses in various fields of applied microbiology.
The application of a binomial distribution to the fractional results of triplicate BIs is only slightly more complicated; the odds of getting positive or negative BI results are not 50–50 because of the natural variability from sample to sample.
As the average number of surviving spores approach zero during a lethality process, the number of spores on individual samples approach a Poisson Distribution which is used to describe probability when the average outcome of an event can be calculated and the results of individual events don’t influence each other.
Currently, many different strategies are used when implementing BIs to validate the decontamination process.
It is advisable that those companies continuing to use single BIs for validation with the expectation of 100% negative growth results allow for a contingent followup test using multiple BIs when an occasional growth positive result is noted.
Appropriate statistical methods may be used to support the acceptance of such rogue results in both primary validation and revalidation programs.” Statistical methods used to interpret BI results require more than one sample per location.
PIC/S recommendations on isolators describe the limitations of using single BIs for validation: “If there is only one BI in each position, and only growth/no growth is established, then the number of survivors is unknown and the size of the possible variation in the process cannot be estimated.” The document describes approaches using single or duplicate BIs in each location.The PIC/S document goes on to describe the approach that has become common place today: “Another possibility is to place three or more BIs at each position in the isolator and put them individually into broth for incubation.If there are any positive broths, the proportion of positive to negative can be used to estimate the number of survivors and thus the log reduction.” The Most Probable Number (MPN) calculation can be used as the basis for estimating the log reduction of spores obtained when using multiple BIs for validation.Complete kill of the BIs was usually expected for process qualification to be considered successful.Isolator operators, however, sometimes experienced difficulties with an occasional positive BI result during the initial qualification or annual requalification.Over time, isolator operators began to realize that these occasional BI issues were not caused by maintenance issues.Tags: Adult Dating, affair dating, sex dating