Effect of Over-The-Counter Drugs on DNA Analysis of Blood Ingested by Medicinal Leeches
Open Access
- Author:
- Patel, Dipesh
- Area of Honors:
- Biology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Reena Roy, Thesis Supervisor
Charles T Anderson, Thesis Honors Advisor - Keywords:
- Drugs
DNA Analysis
Leeches
Aspirin
Ibuprofen
Caffeine
HPLC
DNA Extraction
DNA Quantification - Abstract:
- Painkillers, such as aspirin (acetylsalicylic acid), are used by many to alleviate the pain associated with common ailments. Although the half-life for aspirin is only 2-5 hours it is possible that there may still be sufficient amounts of it in the body post-mortem. Hence, if a medicinal leech, Macrobdella Decora, fed on the blood of a victim or a suspect who had recently ingested an analgesic, it may be possible to simultaneously obtain a drug identification and a complete DNA profile from the human blood extracted from the gut of the leech. The potential for generating a DNA profile depends on whether or not acetylsalicylic acid, ibuprofen, and caffeine inhibit the extraction and quantification of DNA due to their aromaticity. Male human blood was spiked with 1:1 ratio of aspirin, caffeine, and ibuprofen solutions followed by DNA extraction done at 0, 24 and 48 hours. The extracted DNA was then quantified using the PowerPlex® Fusion 6 C system for autosomal study and HPLC was then used to determine the presence of the drugs in the blood samples. A standard amplified product was also examined by capillary electrophoresis and genetic analyzer 3130xl to develop a DNA profile which was used as reference for all future DNA samples which were extracted using this method as all blood used in this experiment was from the same male donor. Blood mixed with the drug was fed to medicinal leeches which were then euthanatized by freezing at 0, 24, and 48 hours after being fed the spiked blood. Human blood from their gut was extracted and analyzed for the presence of aspirin, ibuprofen, and caffeine in order to see a full DNA profile can be generated. Samples were also analyzed with an HPLC to determine the presence of the drugs in the blood samples. The HPLC results concluded that aspirin, ibuprofen, and caffeine were present in substantial quantities in the spiked blood samples collected from the midgut of the leeches. While the leeches did provide a full DNA profile of the male donors, when the collected results were analyzed, the hypothesis that inhibition was caused by acetylsalicylic acid, Ibuprofen, and caffeine was rejected as no inhibition of DNA amplification was noted in both the spiked blood and the blood collected from the midgut of the leeches. Some samples contained higher IPC CT values, but no inhibition was noted upon quantification. Due to this, it can be concluded that acidic and basic over-the-counter drugs containing aromatic groups can be detected in blood samples. These samples can also be used in DNA extraction, quantification, and amplification methods in order to generate a full DNA profile.