Characterization of human DNA in environmental samples.
Forensic Science International, 2008
Authors
- JCJarrod Champagne
- SSStreet IV, W. S.
- MTMary H. Toothman
- KKKaren M. Kester
- BBBonnie L. Brown
- TCTracey Dawson Cruz
Journal
Forensic Science International
Study Design
Addressed Question
analysis of the presence and composition of DNA from environmental dust samples
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
36 samples from different areas
Replicates per Individual and Condition
1
Nucleic Acid
Bodily Origin
Depositor & Contact
Depositor Characteristics
N/A
Criteria for Shedder Status
N/A
Previous Activities
N/A
Contact Scenario
no alteration of sampling site prior to sampling - sampling
Primary Substrate
Primary Substrate Type
surfaces within academic building with low, medium or high human traffic
Primary Substrate Material
Deposit
dust visibility: unapparent, low, medium, high
Delay
N/A
Secondary Substrate
Secondary Substrate Type
N/A
Secondary Substrate Material
N/A
Secondary Substrate Contact
N/A
Further Transfer
N/A
Sampling
Background DNA on Sampled Surface
Sampling Time
direct/delayed
Persistence
N/A
Sampling Method
sterile cotton-tipped swabs moistened with 1.7 ml cell lysis solution
Sampling Area
576 cm^2 area
Laboratory Analysis
Extraction
Mo Bio UltraClean BloodSpin purification kit, final extraction volume: 50 µl
DNA Quantification
total DNA: µQuant MQX200 plate reader, human DNA: Quantifiler Human DNA Quantification kit
Input for Profiling
1-2.5 ng or up to 20 µl of extract
Profiling
AmpFlSTR Profiler Plus PCR amplification kit, MJ BaseStation 51 DNA fragment analyzer, Cartographer v1.2.6sg and ABI Prism 3100 Avant, ABI GeneMapper ID Software v3.2, threshold: 75 rfu
Reference Samples
not available
Profile Interpretation and Mixture Analysis
allele calling following a protocol validated for casework samples and checking by two operators
RNA Data Interpretation
N/A
Results
DNA Quantity
9-28 ng/cm^2 total DNA, 0.2-1.1 pg/cm^2 human DNA
Profile Quality
partial profiles (avg. 0-5 loci/sample) generated from 25/36 samples
Parameter Used for Comparison
total DNA, human DNA (presence and quantity), Genotype recovery (= number of STR loci amplified)
Summary of Results
PCR of human nuclear TAP25 locus and mtDNA locus: human DNA present in almost all samples; high variability in amounts of DNA detected, no clear trends detectable among location, traffic level, total DNA, human DNA or percentage human to total DNA; STR recovery not based on human traffic or visible dust level but dependent on input DNA; high MW loci most often drop-out -> degradation of DNA in the sample; presence of multiple contributors detected; MJ Base Station 51 reveals more peaks/locus
Raised Questions
presence of mtDNA vs. Nuclear DNA <-> age of sample?; implementation of WGA on environmental dust samples in order to improve genotyping results
Cautionary Remarks
additional environmental variables (e.g. light, heat, moisture, cleaning protocols, ventilation systems) not taken into account