Extended Data Fig. 7: Evaluation of NEST performance under polymicrobial contamination across diverse microbial species combinations.
From: One-day rapid sterility test for human-derived biopharmaceuticals
A total of 45 pairwise combinations were generated using 10 microbial species: Staphylococcus epidermidis (ATCC 12228), Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 6538), Streptococcus mitis (ATCC 49456), Pseudomonas aeruginosa (ATCC 9027), Klebsiella pneumoniae (ATCC 13883), Acinetobacter baumannii (ATCC 19606), Enterococcus faecalis (ATCC 29212), Enterococcus faecium (ATCC 19434), and Candida parapsilosis (ATCC 22019). To mimic polymicrobial conditions, MSC solutions were prepared by spiking two microbial species at a concentration of 10 CFU/mL each (total 20 CFU/mL, N = 1), followed by the NEST assay under aerobic conditions. For comparison, single-species samples (10 CFU/mL) were processed in parallel using NEST and served as controls (N = 1). NEST demonstrated 100% detection sensitivity across all tested conditions. Consistent with our previous observations, the detection time for polymicrobial samples was predominantly determined by the faster-growing species. In 44 out of 45 combinations, samples tested positive within one hour of the detection time observed for the faster-growing species. A moderate delay (2.5 h) was observed for the E. faecium/C. parapsilosis combination, potentially due to interspecies growth interference, although contaminant detection remained unaffected.
