In the United States, approximately 100 patients develop fatal sepsis associated with platelet transfusions every year. Current culture methods take 24-48 hours to acquire results, which in turn decrease the shelf life of platelets. Many of the microorganisms that contaminate platelets can replicate easily at room temperature, which is the necessary storage temperature to keep platelets functional. Therefore, there is a need for in-situ quality control assessment of the platelet quality. For this purpose, a real time spectrophotometric technique has been developed. The Spectral Acquisition Processing Detection (SAPD) method, comprised of a UV-vis spectrophotometer and modeling algorithms, is a rapid method that can be performed prior to platelet transfusion to decrease the risk of bacterial infection to patients. The SAPD method has been used to determine changes in cell suspensions, based on size, shape, chemical composition and internal structure. Changes in these cell characteristics can in turn be used to determine microbial contamination, platelet aging and other physiologic changes. Detection limits of this method for platelet suspensions seeded with bacterial contaminants were identified to be less than 100 cfu/ml of sample. Bacterial counts below 1000 cfu/ml are not considered clinically significant. The SAPD method can provide real-time identification of bacterial contamination of platelets affording patients an increased level of safety without causing undue strain on laboratory budgets or personnel while increasing the time frame that platelets can be used by dramatically shortening contaminant detection time.
Recent developments in the characterization of particle dispersions have demonstrated that complementary information on the joint particle property distribution (size-shape-chemical composition) of micron and sub-micron particles is available from multiwavelength spectrophotometric measurements. The UV-VIS transmission spectra of the microorganism suspensions reported herein were recorded using a Hewlett-Packard 8453 diode array spectrometer with an acceptance angle smaller than 2 degrees. To eliminate concentration and particle number effects, the transmission spectra were normalized with the average optical density between 230-900 nm. Experimental results demonstrate that microorganisms at various states of growth give rise to spectral differences that can be used for their identification and classification and that this technology can be used for the characterization of the joint particle property distribution for a large variety of continuous, on-line, and in-situ particle characterization applications. An interpretation model has been developed for the quantitative interpretation of spectral patterns resulting from transmission measurements of microorganism suspensions. The interpretation model is based on light scattering theory and spectral deconvolution techniques and yields the quantitative information necessary to define the probability of the detection and identification of microorganisms. A data base of 54 pathogens has been created and demonstrates that the technology can be used in the field for real-time in-situ monitoring applications.