Genetic individual identification from dried urine spots: A complementary tool to drug monitoring and anti‐doping testing

Abstract

The collection of liquid biological matrices onto paper cards (dried matrix spots [DMS]) is becoming an alternative sampling strategy. The stability over time of molecules of interest for therapeutic, sport drug monitoring, and forensic toxicology on DMS has been recently investigated representing a reliable alternative to conventional analytical techniques. When a tampering of a urine sample in drug monitoring or doping control cases is suspected, it could be relevant to know whether genetic profiles useful for individual identification could be generated from urine samples spotted onto paper (dried urine spot [DUS]). To understand the influence of sex, storage conditions, and time on the quality and quantity of the DNA, five female and ten male urine samples were dispensed onto Whatman 903 paper and sampled after different storage conditions over time, from 1 to 12 weeks. Direct PCR was performed starting from 2‐mm punches collected from each spot amplifying a panel of markers useful for individual identification. The female DUS stored in different conditions produced genetic profiles fully matching the reference samples. The same result was obtained for the male DUS but using urine 30X concentrated by centrifugation instead of the original samples. Our data show that this approach is valid for genetic individual identification of urine samples spotted onto paper cards up to 12 weeks after deposition and could be easily incorporated in anti‐doping or drug screening protocols to help on the suspicion of evidence tampering or to solve questions on the reliability of samples collection.

Advances in testing for sample manipulation in clinical and forensic toxicology - Part A: urine samples

“…Grignani et al evaluated the possibility of using dried urine samples as a source of DNA for personal identification purposes. The chosen approach proved valid for individual genetic identification from dried urine samples stored for up to 12 weeks, which could be helpful in anti-doping or drug screening if tampering is suspected [ 71 ].…”

Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

“…The complication of detecting tampering and manipulation is an inherent conundrum with the attempt of tampering (e.g., by urine substitution) itself being clandestinely done, aiming at remaining undetected. Hence, the use of DNA analyses in support of verifying or falsifying the donorship of a urine sample by a certain athlete has received growing interest, and the possibility to successfully provide DNA profiles from dried urine spots was demonstrated by Grignani et al 99 In a proof‐of‐concept study, urine samples and buccal swabs as comparator specimens were collected from 10 male and 15 female study volunteers, and 50 μl of urine or the pellet formed from 1.5 ml of centrifuged urine was dried on DBS cards. A total of 16 autosomal short tandem repeats (STRs) were used to establish DNA profiles, and although 50 μl spotted from female urine was sufficient for matching test results of urine and buccal swab tests, male urine necessitated the use of concentrated urine particulate.…”