Comparisons of protocols for enhanced DNA profile quality from FTA®-deposited urine samples
Issued Date
2022-12-01
Resource Type
ISSN
18751768
eISSN
1875175X
Scopus ID
2-s2.0-85139673085
Journal Title
Forensic Science International: Genetics Supplement Series
Volume
8
Start Page
133
End Page
135
Rights Holder(s)
SCOPUS
Bibliographic Citation
Forensic Science International: Genetics Supplement Series Vol.8 (2022) , 133-135
Suggested Citation
Srisiri K., Sathirapatya T., Bandhaya A. Comparisons of protocols for enhanced DNA profile quality from FTA®-deposited urine samples. Forensic Science International: Genetics Supplement Series Vol.8 (2022) , 133-135. 135. doi:10.1016/j.fsigss.2022.10.009 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/83523
Title
Comparisons of protocols for enhanced DNA profile quality from FTA®-deposited urine samples
Author(s)
Author's Affiliation
Other Contributor(s)
Abstract
Disputes over the identity of a urine sample donor have been reported, and urine authentication by genetic profiling has helped resolved the cases. However, since genotyping of urine is not always required, many drug-testing laboratories may face sample storage issues. Several studies have investigated the use of FTA® cards as a convenient tool for keeping specimen at room temperature for extended periods of time. However, generating complete STR profile from some FTA®-deposited urine samples remains challenging due to low levels of genetic material content, necessitating amendments to the laboratory's standard protocols. This work therefore aims to evaluate the effects of two DNA template preparation methods, both employing FTA® cards as the storage medium, on the success rates of STR profiling from urine. Specimen from a female volunteer, representing a particularly low-yield sample, was employed. Aliquots of 1 and 2 mL were used as the starting material to evaluate DNA template preparation using the FTA® manufacturer's protocol for disc purification against elution of DNA from the FTA® using Prepfiler™ Forensic DNA Extraction Kit. AmpFℓSTR™ Identifiler™ Plus PCR Amplification Kit was used to amplify the STR markers, and the PCR products were analysed using Applied Biosystems™ 3500xL Genetic Analyzer. The DNA profile qualities were examined in terms of number of loci detected and peak height balance. Comparisons with the profiles obtained from DNA isolated using QIAamp® DNA Micro Kit from 1 and 2 mL of the same batch of urine were also made. The optimised protocol was then tested on urine samples from three male volunteers. The results showed that the purification of FTA® punches according to the manufacturer's protocol enabled full DNA profiles to be obtained from both 1 and 2 mL of urine from all samples tested, including male samples. In contrast, no DNA profile could be generated from the DNA eluted with the Prepfiler™ kit. When compared with the more conventional solid-phase DNA extraction method, the profiles generated from the FTA® punches exhibited similar reproducibility and quality to those from the template isolated by the QIAamp® Kit. This work further demonstrated the feasibility of FTA® cards as a tool for specimen storage and DNA template preparation from small volumes of urine for authentication by STR profiling. Full STR profiles could be generated from sample from both sexes without modification of the PCR conditions or injection time.