Identification of Colored Dyes that are Resistant to Fading on Exposure to Ethylene Oxide; Use with Indicating FTA™ Sample Collection Cards

Abstract

Regulatory Standards and Forensic Communities are expressing an expectation for HID products to be certified as “DNA-free.” Recently, “DNA-free” status was described for HID-related products using ethylene oxide (EtO); this gas reduces the presence of amplifiable DNA and causes minimal interference to downstream HID-analytical methods. During sample collection, indicating cards, for example, Indicating FTA™ (GE Healthcare Life Sciences, UK), are used to collect and store buccal cell DNA. These cards contain a dye which changes color on application of a colorless sample. Generating “DNA-free” indicating cards using EtO should not impact the dyes' ability to indicate sample location or the efficacy of the card in downstream HID-analytical methods. This study was initiated to identify alternative dyes to those currently used with sample indicating collection cards. The most promising, dyes when applied to cellulose papers exhibited a uniform color distribution and excellent sample indicating properties even when mixed with chemicals associated with FTA™. When dyed cellulose papers were exposed to EtO, ultraviolet radiation, elevated temperature, and humidity, negligible fading or discoloration was observed. The presence of these dyes on cellulose papers did not interfere with direct short tandem repeat (STR) profiling. Allelic concordance, first pass success rate, and mean peak heights were comparable to samples applied to Indicating FTA. Biological samples applied to EtO-treated dyed cellulose papers and stored >1 month produced full STR profiles of sufficient quality to allow submission to DNA databases, confirming negligible interference from EtO treatment. These alternative sample indicating dyes resist EtO-mediated fading while fulfilling the Forensic Community's expectation for “DNA-free” with negligible impact on collection card performance.

Keywords

• Ethylene oxide
• forensic DNA analysis
• identification of colored dyes
• Indicating FTA sample collection cards
• short tandem repeat profiling