Abstract:Bacillus subtilis var.niger is an important indicator microbiology of biological protection and sterilization effect evaluation, and its accurate value can ensure the effectiveness and comparability of the evaluation. Spread plate method for bacillus subtilis was method validated, collaborative experimental verified, and uncertainty analyzed. The results showed that spread plate method has a good reproducibility and repeatability, within-laboratory relative standard deviation of repeatability is 10.0%, and inter-laboratory relative standard deviations is 14.1%, when the number of colonies on the plate is between 30CFU and 300CFU. The statistic test showed that the relative expanded uncertainty of spread plate method is 12.0% (k=2). This method can be used for quantitative measurement of bacillus subtilis var. niger spore, which has important significance for the evaluation of protective performance of personnel protective equipment, detection of disinfectants and quality control of sterilization.
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