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Use of Novel Chemistry & Microwave Instrumentation To Improve Body Fluid Assay Sensitivity & Speed While Reducing Costs

NCJ Number
251915
Author(s)
Donald Siegel
Date Published
July 2017
Annotation
Findings and methodology are presented for a research project that developed a proteomic mass spectrometry (MS) assay for blood, saliva, and semen.
Abstract
Current body-fluid testing in common case work requires multiple assay systems. This type of consecutive testing is costly and consumes time and sample. The current project tested and evaluated new technologies and integrated them into current assays in order to make them faster, more accurate, more sensitive, and less expensive. It also examined the simultaneous extraction of protein and DNA from samples, so as to conserve sample use. The research found that microwave digesting significantly reduced protein digestion time from overnight to less than 1 hour, and CPLC (combinatorial ligand peptide chromatography) reduced menstrual blood processing time from days to hours. The researchers are confident that further work, which is described in this report, will improve mixture deconvolution detection levels even more than was achieved with the current assay. The project had four phases: 1) evaluation of CPLC as a means of rapidly preparing complex biological fluids for MS analysis; 2) reduction of trypsin digestion time by evaluating the use of organic solvents and microwave technology; 3) determination of the stability of menstrual blood and vaginal fluid markers over time, evaluation of newly identified microbial vaginal fluid markers, and determination of the effect of forensic reagents Bluestar and luminol on these markers; and 4) determination of the efficacy of simultaneous protein and DNA extraction for use in body fluid and STR testing to reduce sample consumption. 48 figures, 25 tables, and 88 references