The findings and methodology are reported for a research project that showed why loop-mediated isothermal amplification (LAMP) should be considered for a variety of important forensic applications, particularly body fluid identification (bfiD) and Y-screening.
This project tested LAMP as a means of rapidly identifying body fluid, including venous blood, menstrual blood, semen, saliva, and vaginal fluid; and the same approach was applied to male-specific DNA as a potential Y-screen. The use of LAMP enables colorimetric detection by using an array of dyes, which enables the use of inexpensive detection in the form of a smart phone or comparable camera. This project provides evidence that the LAMP method used is as good or better than current presumptive and confirmatory testing. It eliminates human subjectivity in decisionmaking, requires a small footprint, and will be adapted to a 96-well format, tphus indicating the potential for portability and field use. Once optimized, this method should be readily integrated into current forensic casework. Upon being commercialized, it will provide an inexpensive, reliable, alternative to existing bfiD tests. This report describes the project’s sample collection procedures, Y-screen sample preparation and lysis, RNA isolation, DNA quantification, colorimetric loop-mediated isothermal amplification, P30 testing, STR analysis, and the instrument hardware component and operating software. Analytical results are presented for body fluid assay development, body fluid assay testing with mock samples, and Y-screening assay development. Future efforts related to this project are discussed. 34 figures, 8 tables, and 49 references
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