Rapid DNA Maturity Assessment

National Institute of Standards and Technology

Technology Administration, U.S. Department of Commerce

Email: 

Erica.Romsos@nist.gov

Erica L. Romsos1, Sanae Lembirick2, and Peter M. Vallone1

1 U.S. National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899-8314, USA

2 Montgomery College, Rockville, MD 20850, USA

Poster available for download from STRBase: 

http://www.cstl.nist.gov/biotech/strbase/pub_pres/RomsosISFG2015RapidDNA.pdf

P-148

Disclaimer: Points of view in this document are those of the authors and do not necessarily represent the official position or policies

of the U.S. Department of Commerce. Certain commercial equipment, instruments, and materials are identified in order to specify

experimental procedures as completely as possible. In no case does such identification imply a recommendation or endorsement by

NIST, nor does it imply that any of the materials, instruments, or equipment identified are necessarily the best available for the

purpose.

Financial disclosure: This work was supported by funding from the FBI Laboratory and Biometrics Center of Excellence: Forensic

DNA Typing as a Biometric tool.

References:

1. Tan E, Turingan RS, Hogan C, Vasantgadkar S, Palombo L, Schumm JW, et al. Fully integrated, fully automated generation of short 

tandem repeat profiles. Investig. Genet. 4 (2013) 2041-2223.

2. Jovanovich, S., Bogdan, G., Belcinski, R., Buscaino, J., Burgi, D., Butts, E. L. R., et al. Developmental validation of a fully integrated 

sample-to-profile rapid human identification system for processing single-source reference buccal samples. Forensic Sci. Int. Genet. 

16 (2015) 181-194.

3. Hennessy LK, Mehendale N, Chear K, Jovanovich S, Williams S, Park C, et al. Developmental validation of the GlobalFiler express 

kit, a 24-marker STR assay, on the RapidHIT System. Forensic Sci. Int. Genet. 13 (2014) 247-258.

Integration of the extraction, amplification, separation, and detection processes for forensic DNA typing is a challenging goal. Several parallel efforts have been made to integrate the forensic 

workflow and utilize a simple swab in, answer out process within a single platform [1-3]. Of the multiple efforts, two platforms were tested as a part of a rapid DNA maturity assessment in 2014. 

The assessment was conducted with sets of blinded single-source reference samples to gauge the typing success of the current rapid DNA typing technology. Samples were provided to 

participants for testing on the individual rapid platforms, and data was returned to the National Institute of Standards and Technology (NIST) for review and analysis. Both automated and manual 

review of the data sets were conducted to assess the success of typing the CODIS core loci. Genotyping profiles from the multiple platforms, participants, and STR typing chemistry was combined 

into a single analysis to assess the current maturity of Rapid DNA technology. The presented results will focus on genotyping success rate, peak height ratios, and stutter artifacts across two 

platforms and multiple STR kit chemistries.

ANDE (NetBio) RapidHIT 200 (IntegenX)

What is Rapid DNA Typing?

Rapid DNA, or Rapid DNA Analysis, describes the fully automated (hands 

free) process of developing a CODIS Core STR profile from a reference 

sample buccal swab in less than 2 hours. The “swab in – profile out” 

process consists of automated extraction, amplification, separation, 

detection and allele calling without human intervention. 

Integrated Rapid DNA Typing Devices Tested 

As part of the Maturity Assessment

2014 NIST Rapid DNA Maturity Assessment

- Electrophoresis takes place on chip

- Kit = one biochipset

Stored at room temperature

Shelf life ≈ 6 months

- RFID swabs tagged for sample tracking

- Electrophoresis takes place on an 8 

capillary array

- Kit = 4 separate components 

Stored between room temp and 4°C

Shelf life ≈ 6 months at 4°C

- Cotton swabs

GlobalFiler Express loci

≈116 min runtime

(1-7 samples)

PowerPlex 16 loci

≈86 min runtime

(5 samples)

PowerPlex 16 loci

≈90 min runtime

(5 samples)

Rapid DNA Analysis 

vs. Modified Rapid DNA Analysis

Rapid DNA analysis describes the fully automated (hands-free) process 

of developing a CODIS Core STR profile from a known reference sample. 

The “swab in – profile out” process consists of automated extraction, 

amplification, separation, detection and allele calling without human 

intervention. 

Modified Rapid DNA analysis describes the automated (hands-free) 

process of developing a CODIS Core STR profile from a known reference 

sample. This “swab in – profile out” process consists of automated 

extraction, amplification, separation, and detection without human 

intervention but requires manual interpretation and technical review.

NIST provided 20 singlesource reference buccal 

swabs to each participant

RapidHIT 200

PowerPlex 16 

Chemistry

RapidHIT 200

GlobalFiler Express

Chemistry

ANDE

PowerPlex 16

Chemistry

NIST reporting success rate for all 

data/systems combined 

(% successful profiles)

Instrument 

Platforms (2)

Participating 

Laboratories (7)

Independent 

Instruments (11)

NetBio

ANDE

IntegenX

RapidHIT 200

U.S. 

Federal 

State 

Private

5

6

Total Samples 

Tested (280)

100

60

Chemistry

PowerPlex 16

PowerPlex 16

GlobalFiler

Express 120

Data transferred to NIST for analysis

Data transferred to NIST for analysis

Data transferred to NIST 

for analysis

One chemistry and one instrument 

per set of 20 swabs

One chemistry and one instrument 

per set of 20 swabs

One chemistry and one 

instrument per set of 20 

swabs

The purpose of the 2014 NIST Rapid DNA Maturity Assessment was to assess the current status of rapid DNA typing technology for 

the CODIS core loci in support of lab and future external (non-lab-based) Rapid DNA instrument implementation.

Only integrated (swab in – allele detection) instruments capable of genotyping the core CODIS 13 STR markers were eligible for 

this study.

Stutter Percentages

76% 74% 70%

Average Success Rate:

Rapid DNA Analysis

Analysis without human intervention

80% 76% 75%

Average Success Rate:

Modified Rapid DNA Analysis

Analysis involved manual interpretation

Stutter percentages were calculated for all complete profiles 

for the PowerPlex 16 and GlobalFiler Express chemistries. 

Peak Height Ratios

Peak height ratios were calculated for all complete profiles for 

the PowerPlex 16 and GlobalFiler Express chemistries. 

PowerPlex 16 GlobalFiler Express

The PowerPlex 16 data is a 

combination of the data 

generated from both ANDE 

and the RapidHIT 200. 

Additional Resources

1. https://www.fbi.gov/about-us/lab/biometric-analysis/codis/rapid-dna-analysis

2. http://www.swgdam.org/

3. https://www.fbi.gov/about-us/lab/biometric-analysis/codis/summary-of-rapid-dna-addenda-effective-12-1-14

4. https://www.fbi.gov/about-us/lab/biometric-analysis/codis/rapid-dna-addendum-to-qas-final-effective-12-1-2014

5. https://www.fbi.gov/about-us/lab/biometric-analysis/codis/audit-document-for-rapid-dna-qas-addendum-effective-12-1-2014

n=118

n=67

PowerPlex 16

The PowerPlex 16 data is a 

combination of the data 

generated from both ANDE 

and the RapidHIT 200. 

n=118

GlobalFiler Express

n=67

https://www.fbi.gov/about-us/lab/biometric-analysis/codis/rapid-dna-addendum-to-qas-final-effective-12-1-2014

https://www.fbi.gov/about-us/lab/biometric-analysis/codis/rapid-dna-addendum-to-qas-final-effective-12-1-2014

All peak height ratios above 79% Stutter within observed developmental validation range

Genotyping Success

Success: Per Locus

PowerPlex 16

GlobalFiler Express

Percentage of successful genotypes generated per locus. The PowerPlex 16 data is a 

combination of the data generated from both ANDE and the RapidHIT 200. 

Success 

above 80% 

except at Y+ 

and D8S1179

Success 

above 80%

Success was measured by complete and concordant genotypes produced by the integrated rapid DNA devices as compared to 

lab generated correct genotypes

Success rates indicate the average success for each STR locus group genotyped (CODIS 13 loci, PowerPlex 16 loci, new CODIS 

20 loci). The minimum and maximum success rates observed within individual participants within the Maturity Assessment is 

represented by the whiskers of the boxplot above.

STR Loci Genotyped STR Loci Genotyped

Conclusions

Two fully integrated platforms were included in the 2014 Rapid DNA Maturity Assessment. A total of 

11 instruments were tested between 7 laboratories for a total of 280 samples examined. Data for the 

Maturity Assessment was generated from October-December 2014 and returned to NIST for data 

analysis. Updates to instrumentation and software may have taken place since this Maturity 

Assessment was completed. Observed success for the CODIS 13 Core Loci ranged from 76% for 

Rapid DNA Analysis to 80% with Modified Rapid DNA Analysis. 

Results can be located: http://www.nist.gov/mml/bmd/genetics/dna_biometrics.cfm