Optimisation of DNA recovery and analysis of urine samples stored on FTA® card

Abstract

Denial of ownership of urine samples submitted by athletes for doping control has led to an increasing need to incorporate DNA analysis into testing protocols. As there are often large volumes of samples to process, a streamlined sample collection and analysis protocol suitable for urine samples that is quick, reliable, and economical would be highly advantageous. FTA® cards have been employed extensively for preparing and preserving genetic material for long periods of time at room temperature. This study reports the possibility of using FTA® cards to store urinary DNA, which can be used directly as templates in STR analysis for human identification. Urine samples from six male and female volunteers were collected. The sample preparation steps for harvesting cells from urine, such as minimum urine volume required and centrifugal speed, were optimised, and suitable sizes of the FTA® card punches were determined. The results showed that regardless of the sample donor’s sex, a complete DNA profile could be reproducibly generated from a single 2.0-mm punch of the 3-mL urine samples that had been deposited on FTA® cards using the standard PCR conditions for AmpFLSTR® Identifiler® Plus. The developed protocol could become an integral part of a guideline for authentication of urine samples submitted for doping control or forensic toxicological analysis, where cost per sample and storage space are factors of consideration.