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Direct inoculation or membrane filtration, 14d at 20-25°C & 30-35°C
DNA extraction, 16S/18S rRNA amplification and sequencing
Recovery test with B.subtilis spike on products
Direct LAL test on water sample
Membrane filtration, R2A agar incubation 7d at 20-25°C
Enrichment in LST broth, confirmation on EMB/MacConkey agar
Growth promotion with 5 strains on TSA plates
Biochemical profiling from pure colonies
Crystal violet/iodine/safranin staining of colonies
Membrane filtration, TSA plate count, 3d at 30-35°C
Decontamination, concentration, culture on Löwenstein-Jensen
Method suitability test with E.coli ATCC 8739
Bacteriostasis/fungistasis test with 6 challenge organisms
Protein fingerprinting by mass spectrometry from colonies
Selective enrichment and plating for target organisms
Spore enumeration by heat shock and plate count
Recovery efficiency + method suitability + 2 ID of dominants
Interference test at 3 dilutions with spike recovery
Method suitability study for product-specific sterility testing
Selective enrichment at 42°C, microaerophilic conditions
Heat shock, serial dilution, TSA plate count
Extraction/rinse, filtration, TSA 3-5d & SDA 5-7d incubation
Screening at multiple dilutions to find valid test range
Direct incubation in growth media, 7d at optimal temperature
Acid/heat treatment, BCYE agar culture, 10d at 35°C
Growth promotion test with C.albicans, A.brasiliensis on SDA
Extraction, filtration, TSA incubation 3-5d at 30-35°C, aerobic conditions
Anaerobic incubation on TSA or FTM media, 5-7 days
Kinetic chromogenic LAL assay, 0.005-5.0 EU/ml range
Spore suspension filtration and culture incubation 7d at optimal temperature
Challenge test with 5 organisms, log reduction monitoring