An evaluation of the RapidHIT™ ID system for hair roots stained with Diamond™ Nucleic Acid Dye

Abstract

The RapidHIT™ ID (RHID) system was evaluated for its suitability in processing a single hair root to obtain informative DNA profiles. Hair samples were assessed for nuclear DNA prior to DNA analysis using Diamond™ Nucleic Acid Dye (DD) and real-time Extended Depth of Field (EDF) imaging to visualise and count nuclei if present. Hairs were viewed under an Optico N300F LED Fluorescent Microscope and imaged using a MIchrome 5 Pro camera. Hair roots were processed through both the ACE GlobalFiler™ Express sample cartridge and the RapidINTEL™ sample cartridge. A total of 44 hairs including shed hairs (9) and plucked hairs (35) from 8 donors were evaluated in this study. The processing of hairs using the RHID system required the modification of a standard swab that allowed for hairs to be easily collected and placed into the cartridge but also allowed for the re-collection of hair roots post RHID analysis (for potential standard DNA workflow). 90% of plucked hairs with a high nuclei count (>100) resulted in a high partial or full DNA profile, with the remaining 10% resulting in a low partial profile. 44% of shed hairs resulted in a low partial profile, with the remaining hairs resulting in a null profile. This study demonstrated that the RHID system could successfully obtain a DNA profile from a single hair root with nuclei present post-DD staining. According to these results, it is suggested that when dealing with hairs containing fewer than 50 nuclei, using the RapidINTEL™ cartridge can enhance allele recovery.