2018 Rapid DNA Maturity Assessment

Erica Romsos & Peter Vallone

Applied Genetics Group, NIST

November 7, 2018
Steps in Forensic DNA Analysis

Sample Collection

Blood Stain Buccal swab

1-2 day process (a minimum of ~8 hours) with current 

laboratory procedures and technology

DNA 

Extraction

1.5 

hours

DNA 

Quantitation

1.5 

hours

Multiplex PCR 

Amplification

3.5 hours

Interpretation of Results

STR Typing

DNA separation and sizing

1.5 

hours

Collection

Extraction

Quantitation

Multiplex PCR

Steps Involved

Interpretation 

of Results

STR Typing
Commercial RDNA Instruments

• RapidHIT 200

• PowerPlex 16HS

• Globalfiler

• RapidHIT ID

• Globalfiler

IntegenX ANDE

3-7 Samples

2 hour run time

1 Sample

90 minute run time

5 Samples

86 minute run time

• ANDE/DNAScan

• PowerPlex 16

• ANDE

• FlexPlex (27 loci)
Rapid DNA Instrument Testing

2010: Apollo 200 Testing

(136 samples)

09/2012: First prototypes delivered 

to NIST (DHS Instruments)

2013: NIST Interlab

Study (350 samples)

2014: NIST Maturity 

Assessment (280 samples)

2012-2015: Ongoing communication with Developers

2015: NIST Participates in 

IXI PP16 Dev. Validation 

(100 samples)

2016: NIST Participates in 

ANDE PP16 Dev. Validation 

(150 samples)

2017: NIST Participates in 

ANDE FlexPlex Dev Validation 

(250 samples)

2016: NIST Participates in 

testing ParaDNA

2018: NIST Rapid DNA 

Maturity Assessment 

(240 samples)
NIST: Rapid DNA Maturity Assessments

• Collection and distribution of samples to all participating laboratories

• 2013: 3 labs, 150 samples

• 2014: 7 labs, 280 samples

• 2018: 9 participants, 240 samples

• Coordination of all testing sites to include return of all data to NIST for 

analysis and review 

• Includes IRB approval for swab collection and MTA for distribution

• Analysis and compilation of all data

• Summary of results presented across multiple meetings within the 

forensic and biometric communities
2013 Rapid DNA Maturity Assessment

• Summer 2013 we felt confident that we could carry out an interlab

assessment of the R-DNA prototypes

• Data was collected and analyzed in August 2013

• All instruments had the same version of software and scripts for testing

• Three federal testing sites (7 individual instruments)

• 50 single source reference buccal swabs were provided by NIST for each 

instrument tested

• 350 single source reference buccal swabs tested

• Success defined as the automated calling of the 13 CODIS core STR loci

• Overall success = 88.3%
2014 Rapid DNA Maturity Assessment

• Fall 2014 assessment of the current status of rapid DNA typing 

technology for the CODIS Core Loci

• In support of lab and future booking station Rapid DNA implementation

• Many modifications to both hardware and software were made 

between 2013 and 2014

• 7 participating laboratories (11 independent instruments)

• 20 single source reference buccal swabs tested

• 280 single source reference buccal swabs tested

• Success defined as automated (lights out) calling of the 13 CODIS core loci

• Overall Success = 76.1% 
2018 Rapid DNA Maturity Assessment

• Goal: To measure the status of rapid DNA typing technology 

for the 20 CODIS core loci in support of booking station 

Rapid DNA implementation

• Rapid DNA instruments capable of genotyping the 20 CODIS 

core loci were eligible for participation

• 20 single source reference buccal swabs were distributed to 

participating laboratories
NIST provided 20

reference buccal 

swabs to each 

participant

Participant Runs 

RH-ID GFE

Participant Runs 

RH200 GFE

Participant Runs 

ANDE 6C FlexPlex

NIST reports 

CODIS 20 success 

rate for all data 

combined 

(% success)

Data transferred back to NIST via electronic format for 

analysis

Participants are provided one set of 20 buccal swabs per 

instrument with the possibility to request additional sets for 

additional instruments

2018 Rapid DNA Maturity Assessment Study Outline
Timeline of 2018 Maturity Assessment

February 2018-Present: Buccal samples collected at NIST and stored 

at RT (>900 swabs have been collected from >50 unique individuals)

May 2018: Official call for Participants in the 2018 Rapid DNA 

Maturity Assessment for the CODIS core 20 loci

July 2018: Samples shipped to participants 

August 2018: Data returned to NIST for analysis
Instrument 

Platforms

Participants

ANDE 6C

IntegenX RapidHIT 200

Federal 

State

Total Samples 

Tested

100

60

Chemistry

FlexPlex

GlobalFiler

Express

80

20 Swabs provided 

Police

Vendor

IntegenX RapidHIT ID

GlobalFiler

Express

Analysis 

Method

Rapid DNA 

Analysis 

Modified 

Rapid DNA 

Analysis

Modified 

Rapid DNA 

Analysis

Independent 

Instruments

5

3

4

9 Participants 3 Platforms 12 Instruments 240 Samples
Success Metrics

• Success was measured by complete and concordant genotypes 

produced for the 20 CODIS core loci

• Allele calls by the integrated rapid DNA devices were compared to lab 

generated profiles for concordance 

• Fusion 6C, PP21, GFE on a 3500xL

• Two interpretation parameters were implemented 

• Rapid DNA Analysis: Without human intervention

• Modified Rapid DNA Analysis: Expert interpretation and analysis of 

electropherogram https://www.fbi.gov/file-repository/addendum-to-qas-for-rapid-dna.pdf/view
Rapid DNA Analysis-ANDE

Automated (lights-out) analysis without human intervention

To date, ANDE 6C is the only rapid 

DNA system to be NDIS approved 

for automated rapid DNA analysis

https://www.fbi.gov/services/laboratory/biometric-analysis/codis/rapid-dna
Average Success: 85%

n=85
Modified Analysis Parameters

• Samples were manually reviewed (n=240)

• ANDE profiles were analyzed in GeneMapper IDX v1.5 

• RapidHIT profiles were reviewed in GeneMarker HID v2.8.2

• Heterozygote balance filter set at 0.25

• After manually interpreting a profile (PHR >0.25, low AT, recovery of 

“lost” data), concordance was checked against the laboratory 

generated reference profile 

• Success was determined by complete and concordant profiles for the 

20 CODIS core loci
Average Success: 90 %

n=217
Unrecovered Samples 

n=23

Instrument Related 

Unknown

n=9

Data 

Transfer 

Failure

n=2

Partial Profile

n=10

Single 

Locus 

Dropout

n=2
Unrecovered Samples 

n=23

Instrument Related 

Unknown

n=9

Data 

Transfer 

Failure

n=2

Partial Profile

n=10

Single 

Locus 

Dropout

n=2

No data was transferred from the 

instrument to analyze
Unrecovered Samples 

n=23

Instrument Related 

Unknown

n=9

Data 

Transfer 

Failure

n=2

Partial Profile

n=10

Single 

Locus 

Dropout

n=2
Unrecovered Samples 

n=23

Instrument 

Related 

Unknown

n=9

Data 

Transfer 

Failure

n=2

Partial Profile

n=10

Single 

Locus 

Dropout

n=2
Summary of Success

• Automated success rate of 85% (n=100)

• ANDE 6C is the only NDIS approved 

instrument for automated analysis

• xml files for the unsuccessful profiles 

were not generated by the 

instrument (15%)

• Modified analysis success rate of 90%

• Manual interpretation of all samples 

(n=240 samples)

• Increase of success for manually 

reviewed profiles
Additional Metrics Analyzed

Base pair sizing precision

Heterozygote Balance

n=217 successful profiles

64%
n=19 ladders for ANDE

n=8 ladders for RH 200 Precision-Base Pair Sizing
Heterozygote Balance

95% of the samples 

were greater than 52% 

0.52
95% of the samples 

were greater than 63% 

0.63

Heterozygote Balance
Heterozygote Balance

95% of the samples 

were greater than 52% 

0.52
Heterozygote Balance

95% of the samples 

were greater than 59% 

0.59

n=217 successful profiles
Maturity Assessment Summary

• 12 instruments tested across 9 laboratories

• Total of 240 samples examined

• 85% success rate for the CODIS 20 using Rapid DNA Analysis

• 90% success rate for the CODIS 20 using Modified Rapid DNA 

Analysis

• Success ranged from 60% to 100%

• Precision was below 0.17 bp on for both ANDE 6C and 

RapidHIT 200

• Combined heterozygote balance (all three instruments) was 

above 59%
Acknowledgements

Contact Information 

erica.romsos@nist.gov

NIST – Applied Genetics Group

Peter Vallone

Steven Lund

Funding

FBI Biometrics Center of Excellence: Forensic 

DNA Typing as a Biometric tool.

Thank you to our participants

• ANDE

• Arizona Department of Public Safety

• Bensalem Police Department

• Federal Bureau of Investigation Laboratory

• Louisiana State Police Crime Laboratory

• Miami Beach Police Department

• Miami Dade Police Department

• NIST (DHS instruments, run at SNA Intl.)

• U.S. Army Criminal Investigation Laboratory