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At the end of the incubation time (10 days, 20 mL), the negative bottles were also tested by culturing 100 μL of the fluid into the blood, MacConkey, and Brucella agar and incubated under aerobic and anaerobic conditions for 48 h at 37°C.
After incubation, positive as well as negative bottles were subcultured on Sabouraud agar, CHROMAgar Candida, and Columbia agar.
Another approach could have been to perform terminal cultures from the negative bottles in those isolates that were missed by the Bactec9240 system as well as the false positive signals, however this was not part of the study design.
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We did not test identification accuracy in BACTEC-negative bottles, as PNA-FISH requires an organism concentration of at least 10 CFU/mL for detection.
All VRE were cultured from one bottle from a CVC with one or more negative culture bottles from simultaneous peripheral blood cultures in all patients.
Open symbols indicate average of replicate uninoculated negative control bottles.
We observed that some negative Anaerobic bottles (11 %) were positive in subcultures, of which a high proportion was C. glabrata (12 out of 19; 63 %).
Negative blood culture bottles were incubated for seven days before being reporting negative.
Material from inoculated, previously negative blood cultures bottles and henceforth called blood culture material.
This protocol was standardized by testing negative blood culture bottles and C. albicans-spiked blood culture as negative and positive controls, respectively (data not shown).
The reproducibility of the assay was 100% as checked by a pilot study testing negative blood culture bottles spiked with cultured Candida spp. cells (data not shown).
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