While DNA profiling makes it possible to individualize biological stains, the identification of the stain itself can present forensic serologists with a significant challenge. Current antibody- and enzyme activity-based assays used by forensic practitioners for biological stain identification yield only presumptive results. Positive results with non-target body fluids or cross-reactivity with non-human sources has also been well documented. Some tests can consume unacceptable quantities of precious evidence while for some body fluids (vaginal fluid and menstrual blood) there are simply no available tests at all.
This research presented here aims to develop and rigorously test a fast, accurate, and sensitive multiplex assay for the simultaneous identification of saliva, semen, urine, peripheral blood, menstrual blood, and vaginal secretions. This research is based on the following three research phases:
1. Biomarker Identification – Utilizing multidimensional protein separation technologies, bioinformatics tool, and tandem mass spectrometry a database of fluid-specific candidate markers will be developed for each body fluid.
2. Biomarker Verification – Using the most promising candidates from Phase 1 a targeted multiplex assay will be developed on a Quadruple Time-of-Flight mass spectrometer to verify the specificity of these candidate biomarkers with single source laboratory samples as well as single and mixed source casework type samples.
3. Prototype Validation – Development and testing of a case working laboratory prototype assay on a triple quadrupole mass spectrometer using single and mixed source casework-type samples.
This work has the potential to significantly improve the accuracy and sensitivity of forensic serological testing. It will provide practitioners with greatly improved tests for saliva and seminal fluid while also enabling the identification of vaginal secretions for which there are currently no accurate tests. The multiplex design will eliminate the need to perform separate tests on an unknown stain. In short, the successful completion and implementation of this research will provide the forensic community with a powerful tool to aid in criminal investigations.
","abstract_html":"<p>While DNA profiling makes it possible to individualize biological stains, the identification of the stain itself can present forensic serologists with a significant challenge. Current antibody- and enzyme activity-based assays used by forensic practitioners for biological stain identification yield only presumptive results. Positive results with non-target body fluids or cross-reactivity with non-human sources has also been well documented. Some tests can consume unacceptable quantities of precious evidence while for some body fluids (vaginal fluid and menstrual blood) there are simply no available tests at all.</p> <p>This research presented here aims to develop and rigorously test a fast, accurate, and sensitive multiplex assay for the simultaneous identification of saliva, semen, urine, peripheral blood, menstrual blood, and vaginal secretions. This research is based on the following three research phases:</p> <p>1. Biomarker Identification – Utilizing multidimensional protein separation technologies, bioinformatics tool, and tandem mass spectrometry a database of fluid-specific candidate markers will be developed for each body fluid.</p> <p>2. Biomarker Verification – Using the most promising candidates from Phase 1 a targeted multiplex assay will be developed on a Quadruple Time-of-Flight mass spectrometer to verify the specificity of these candidate biomarkers with single source laboratory samples as well as single and mixed source casework type samples.</p> <p>3. Prototype Validation – Development and testing of a case working laboratory prototype assay on a triple quadrupole mass spectrometer using single and mixed source casework-type samples.</p> <p>This work has the potential to significantly improve the accuracy and sensitivity of forensic serological testing. It will provide practitioners with greatly improved tests for saliva and seminal fluid while also enabling the identification of vaginal secretions for which there are currently no accurate tests. The multiplex design will eliminate the need to perform separate tests on an unknown stain. In short, the successful completion and implementation of this research will provide the forensic community with a powerful tool to aid in criminal investigations.</p>","abstract_has_math":false,"creators":["Legg, Kevin M."],"institution":null,"degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Phillip B. Danielson, Ph.D.","Joseph Angleson","James Fogleman","Joyce S. Sterling"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-11-01T07:00:00Z","date_published":"2014-11-01T07:00:00Z","updated_at":"2026-07-24T02:02:11Z","subjects":["Forensics","Mass spectrometry","Proteomics","Serology","Biology","Life Sciences"],"languages":["en"],"rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.du.edu/etd/360","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Phillip B. Danielson, Ph.D.","Joseph Angleson","James Fogleman","Joyce S. Sterling"]},{"key":"dc:creator","label":"Author","values":["Legg, Kevin M."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2001-01-01T08:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Forensics","Mass spectrometry","Proteomics","Serology","Biology","Life Sciences"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright is held by the author. User is responsible for all copyright compliance.
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This research presented here aims to develop and rigorously test a fast, accurate, and sensitive multiplex assay for the simultaneous identification of saliva, semen, urine, peripheral blood, menstrual blood, and vaginal secretions. This research is based on the following three research phases:
1. Biomarker Identification – Utilizing multidimensional protein separation technologies, bioinformatics tool, and tandem mass spectrometry a database of fluid-specific candidate markers will be developed for each body fluid.
2. Biomarker Verification – Using the most promising candidates from Phase 1 a targeted multiplex assay will be developed on a Quadruple Time-of-Flight mass spectrometer to verify the specificity of these candidate biomarkers with single source laboratory samples as well as single and mixed source casework type samples.
3. Prototype Validation – Development and testing of a case working laboratory prototype assay on a triple quadrupole mass spectrometer using single and mixed source casework-type samples.
This work has the potential to significantly improve the accuracy and sensitivity of forensic serological testing. It will provide practitioners with greatly improved tests for saliva and seminal fluid while also enabling the identification of vaginal secretions for which there are currently no accurate tests. The multiplex design will eliminate the need to perform separate tests on an unknown stain. In short, the successful completion and implementation of this research will provide the forensic community with a powerful tool to aid in criminal investigations.
"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Development and Testing of a Rapid Multiplex Assay for the Identification of Biological Stains"]}]}],"canonical_facts":{"dc:contributor":["Phillip B. Danielson, Ph.D.","Joseph Angleson","James Fogleman","Joyce S. Sterling"],"dc:creator":["Legg, Kevin M."],"dc:date.available":["2001-01-01T08:00:00Z"],"dc:description.abstract":["While DNA profiling makes it possible to individualize biological stains, the identification of the stain itself can present forensic serologists with a significant challenge. Current antibody- and enzyme activity-based assays used by forensic practitioners for biological stain identification yield only presumptive results. Positive results with non-target body fluids or cross-reactivity with non-human sources has also been well documented. Some tests can consume unacceptable quantities of precious evidence while for some body fluids (vaginal fluid and menstrual blood) there are simply no available tests at all.
This research presented here aims to develop and rigorously test a fast, accurate, and sensitive multiplex assay for the simultaneous identification of saliva, semen, urine, peripheral blood, menstrual blood, and vaginal secretions. This research is based on the following three research phases:
1. Biomarker Identification – Utilizing multidimensional protein separation technologies, bioinformatics tool, and tandem mass spectrometry a database of fluid-specific candidate markers will be developed for each body fluid.
2. Biomarker Verification – Using the most promising candidates from Phase 1 a targeted multiplex assay will be developed on a Quadruple Time-of-Flight mass spectrometer to verify the specificity of these candidate biomarkers with single source laboratory samples as well as single and mixed source casework type samples.
3. Prototype Validation – Development and testing of a case working laboratory prototype assay on a triple quadrupole mass spectrometer using single and mixed source casework-type samples.
This work has the potential to significantly improve the accuracy and sensitivity of forensic serological testing. It will provide practitioners with greatly improved tests for saliva and seminal fluid while also enabling the identification of vaginal secretions for which there are currently no accurate tests. The multiplex design will eliminate the need to perform separate tests on an unknown stain. In short, the successful completion and implementation of this research will provide the forensic community with a powerful tool to aid in criminal investigations.
"],"dc:format":["application/pdf"],"dc:identifier":["https://digitalcommons.du.edu/etd/360"],"dc:language":["en"],"dc:rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"dc:subject":["Forensics","Mass spectrometry","Proteomics","Serology","Biology","Life Sciences"],"dc:title":["Development and Testing of a Rapid Multiplex Assay for the Identification of Biological Stains"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."]},"updated_at":"2026-07-24T02:02:11Z"}