Direct detection of the bacterial stress response in intact samples of platelets by differential impedance

Abstract

We have previously described a new rapid approach that relies on monitoring intentionally stressed bacteria in contaminated platelet concentrates (PCs). This earlier work included human cell lysis with Triton X-100 and filtration as steps in the sample preparation. This study was undertaken to develop an improved and time-saving protocol that enables direct bacterial detection in PCs without lysis and filtration. Apheresis- or whole blood-derived PCs were spiked with 17 model bacteria and tested at final concentrations from 10(3) to 10(6) colony-forming units (CFUs)/mL. The contaminated PCs were treated with a chemical compound that induces a stress response in bacteria and monitored using differential impedance sensing to detect and record subtle changes in the dielectric permittivities of the contaminated platelet (PLT) samples. No measurable responses from sterile PLT samples were observed during exposure to the compounds used as stressors. In contrast, distinct response profiles were obtained without exception for all 17 bacterial species for all bacterial concentrations tested. Bacterial presence was established within 5 to 10 minutes for high inocula (10(6) and 10(5) CFUs/mL) while low inocula (10(4) and 10(3) CFUs/mL) were usually detectable within 20 minutes. The entire testing process routinely took less than 30 minutes from the point of sampling to the time that the final results are available. The results described here demonstrate that monitoring the development of stress in bacteria is a fast and simple way to detect 10(3) CFUs/mL or more bacteria in complex cellular blood products such as PCs.