Evaluation of a rapid DNA process with the RapidHIT® ID system using a specialized cartridge for extracted and quantified human DNA

Abstract

The turn-around time of urgent crime scene DNA samples is often far longer than desired by law enforcement. Crime scene DNA sample processing involves both complex and routine processing steps. Simplification and integration of the routine steps would dramatically improve turn-around time and reduce the risk of operator contamination. Routine DNA extraction and quantitation is readily available. However, PCR amplification and electrophoretic analysis remain largely manual. Rapid DNA Analysis is a hands-free "swab in - profile out" process which consists of automated DNA extraction, amplification, separation, detection, and allele calling without human intervention. RapidHIT 200 and RapidHIT ID are rapid DNA systems developed by IntegenX (Pleasanton, CA) and validated for the use of buccal swabs. A new generation of the RapidHIT sample cartridge for RapidHIT ID has been designed and tested which allows the loading of extracted and quantified DNA. RapidHIT EXT sample cartridge allows a user to generate a forensic DNA profile from less than 250 pg of extracted and quantified DNA in less than 90 min with less than one-minute hands-on time. Once the sample is loaded in the RapidHIT EXT sample cartridge, a DNA profile is produced after amplification, detection and automated data analysis. We report on sensitivity, reproducibility, concordance, DNA mixtures and carryover for EXT sample cartridges pre-loaded with GlobalFiler Express and AmpFLSTR NGM SElect™ Express, (Thermo Fisher Scientific, Waltham, MA) STR chemistries. Purified and quantified DNA from mock crime scene samples were used to demonstrate the utility of these cartridges in an established forensic laboratory.

Recent trends and developments in forensic DNA extraction

“…As such, RapidHIT 200 System is more catered to users in forensic laboratories. To cater to low throughput needs in a decentralized environments such as police stations or border crossing stations (Salceda et al, 2017), Thermo Fisher also launched the Applied Biosystems RapidHIT ID System that generates STR profiles from one reference sample at a time within 90 min, with either the ACE or EXT microfluidic cartridge, both of which are single‐use (Buscaino et al, 2018; Salceda et al, 2017). The RapidHIT ID with the ACE cartridge has been validated to process reference buccal samples (Amick & Swiger, 2019; Salceda et al, 2017; Shackleton, Gray, Ives, Malsom, & Vanhinsbergh, 2019; Wiley et al, 2017).…”

“…The RapidHIT ID with the ACE cartridge has been validated to process reference buccal samples (Amick & Swiger, 2019; Salceda et al, 2017; Shackleton, Gray, Ives, Malsom, & Vanhinsbergh, 2019; Wiley et al, 2017). DNA extraction is performed in the swab chamber of the cartridge (Buscaino et al, 2018; Salceda et al, 2017; Wiley et al, 2017). The sample in the ACE cartridge is extracted based on the Prep‐ n ‐Go buffer chemistry (ThermoFisher Scientific).…”

Autosomal STR Profiling and Databanking in Malaysia: Current Status and Future Prospects

“…Several companies have developed rapid DNA test machines (e.g., ANDE™ Rapid DNA Analysis System developed by Accelerated Nuclear DNA Equipment, and Applied Biosystems RapidHIT ® ID system by Thermo Fisher Scientific Inc., Waltham, MA, USA) where DNA profiles can be generated in a much shorter time than what is possible using the conventional profiling methodology. In addition, these new DNA profiling platforms are fully automated, and only require a sample injection from swabs to produce STR profiles ready for matching against those already stored in the national DNA databank [ 53 , 54 ]. This means that DNA profiling can now be performed outside of accredited laboratories and by operatives (e.g., police officers or civilian employees) that have only received basic training on the instruments.…”

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

“…These systems are microfluidic based and include the RapidHIT ® ID (Thermo Fisher Scientific, USA) and ANDE™ Rapid DNA Analysis system (ANDE, USA) [14]. Portable rapid systems are designed to be used as deployed capabilities and at non-laboratory settings such as police custody suites, and have mainly been evaluated for reference sample collection [15,16].…”