Direct PCR of bloodstains collected from deceased individuals for identification purposes
| Autor: | Dadna Hartman, Zoe Bowman, Valerie Chahin Atallah |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
First pass
Veterinary medicine 010401 analytical chemistry Sample processing Disaster victim identification Biology 01 natural sciences Dna identification DNA extraction 0104 chemical sciences Pathology and Forensic Medicine law.invention 03 medical and health sciences 0302 clinical medicine law Genetics 030216 legal & forensic medicine Polymerase chain reaction |
| Zdroj: | Forensic Science International: Genetics Supplement Series. 7:406-407 |
| ISSN: | 1875-1768 |
| DOI: | 10.1016/j.fsigss.2019.10.030 |
| Popis: | Direct PCR provides advantages over traditional DNA processing by removing the DNA extraction and quantification steps, thus reducing sample processing time. Such advantages are invaluable in disaster victim identification (DVI) and other cases where DNA identification of a deceased is required. Where possible we collected bloodstains on Copan NUCLEIC-CARD™ in cases where DNA identification of a deceased was required. An attempt was made to develop a direct PCR processing pipeline for bloodstains collected from deceased individuals. Initial testing with GlobalFiler™ Express using the manufacturer recommended 1.2 mm sample punches showed significant inhibition in many samples, with 20 (77%) of the 26 samples tested resulting in full profiles. This first pass success rate is insufficient for our purposes. Another concern were the unusually high baseline artifacts seen in relative proportion to the peak heights obtained. The GlobalFiler™ casework kit was also trialled with 1.2 mm sample punches which demonstrated a cleaner baseline but increased inhibition. To reduce the amount of inhibition, the amount of sample added to the PCR reaction was halved which resulted in four of the previously unsuccessful samples producing a full profile using GlobalFiler™ Express as well as less inhibition effects in other samples tested. GlobalFiler™ casework was also successful with a halved input however more inhibition was observed when compared to GlobalFiler™ Express. The two remaining unsuccessful samples that failed to produce any results had, however, previously given results with traditional profiling. Pre-amplification dilution of samples had some success in producing profiles from these samples. |
| Databáze: | OpenAIRE |
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