SUPA Forensic Conference11-21-14

Speaker:

Michael Marciano

Senior Scientist

Professional Background

 THE DNA PARADIGM

Forensic and National Security Sciences Institute

College of Arts and Sciences  Syracuse Universityppt developed by JoAnne Pawlowski

• Research and Teaching Interests• M.S. Forensic Molecular Biology, University at Albany

The intersection of genetic identity, DNA based forensic science and issues pertaining to national security. Research thrust areas in molecular biology and genetics addressing applied scientific questions to the exploitation of biologicals to aid in tagging, tracking locating targets of interest, DNA based geolocation. Future teaching interests lie in forensic DNA analysis with focuses on current practices, innovative applied research, legal/ethical matters and public speaking and communication.

• ‘My background as a practicing Forensic Scientist at the Onondaga County Center for Forensic Sciences has provided a foundation in field based applications of forensic techniques that bridge the gap between practitioners and the research community. In this time I was recognized in federal and state courts as an expert in DNA analysis. I joined the SRC Bioforensics group in pursuit of new challenges in applied research and development relating to national security and global threats. With Dr. David Knaebel, we engaged both the intelligence and law enforcement communities through our innovative biological solutions to complex problem sets. As a new member of the Syracuse University Forensic and National Security Sciences Institute I look forward to interacting with faculty, staff and students through collaborations, research and classroom environments.’

• Michael Marciano, senior scientist

• Forensic and National Security Sciences Institute

• College of Arts and Sciences  Syracuse University

How DNA effects the other forensic sciencesBackground

Define forensicWhat is DNA ?Beyond markers

Forensic DNA analyst: testify in court and analysis of body fluids

Using DNA to geosource

Locard Exchange Principle-basis for evidence left and takenNo perfect crimeBUT limits based on current sciences- some quite sophisticatedTech today applied to cases years agoInnocence Project examplesForensic in FS means adjudication; application to court or judicial system or problems

CSI effect does exist- demands from court /expectationsTherefore unreasonable amount of evidence requestedPandering to jury with high expectationsLab services involves many units within science principlesAll may be expected to deliver

Forensic labs and accreditation- Nassau County lab closedDNA raised bar of expectations for other sciencesAccreditation-labs responsible for scientific analysis and testimonyDemandingLab accreditation boards - Pre approval of techniques for courtScientific working groups on DNA analysis methodsSame for firearms, toolmarks, drugs, hair, blood spatter etc.Oversight groups are federal.. Working groups now dissolvedWritten into law that these organizations have to be present

TIMELINE OF DNA and Forensic Science

1868 Miescher “discovers” DNA 1953 Watson and Crick report double helix structure 1977 First human gene cloned1985 Jeffreys reports VNTR DNA sequences1985 First report of PCR method 1986 Jeffreys uses DNA to solve first murder case (Pitchfork case) 1987 First conviction on DNA evidence (Andrews case) 1991 STRs first reported-1998 FBI starts CODIS database 2005 2.5 million DNA “fingerprints” in CODIS

Develop PCR to identify individualFirst to testify in court as criminal caseRFLP technology until OJ case

1988 tech group was developed and NY v Castro was decision that required standardization of laboratory testing procedures in fs

FBI established DNA analysis unitOJ-Start of rapid evolution of standardization of DNA analysis

Set standard how NOT to testify as in OJ case.Bruce Weir, Expert.. Google that name Standard stats to be used.. And 13 core loci 1990 CODIS established

DNA applications and limitations and CODIS:

Loci expanded to 24.. More discrimination Exoneration or exclusion of a suspect.. Not only inclusionCorroboration of testimony from witnessPaternityIdentification of bodies after disasters

First DNA cases; Saratoga NY Sexual assault. Pre CODISDescribed assailant.. Saw lineup.. Convicted went to prison.Later.. Year or two.. DNA analysis on rape kitNot the guilty man-.guys looked alike

Will study Thomson and Cotton case

Eye witness testimony still evidence problem

When no comparison in CODIS ..problemCODIS: still lackingStates differ on who may be typed (not like fp and all arrested)Profile into CODIS must be typed in 3 x.. Not cut and pastedGenes into LODIS-localThen SODIS- state, then NODIS- National-.those are all what is CODIS now

Therefore, Combined- CODIS

List sources of DNA… all possible.Identify body fluidsWhat to pick out in room to analyze..The cost and time for each sample is large.. Must be discriminatedWhat is touched.. Or put to mouth..Alternate light sources used: diff wavelengths.. What fluoresces?Body fluids.. Proteins.. Fatty acids will glowSingle human cell has 6.6 pg of DNA1 ng of Human DNA = 152 diploid cellsCan get DNA from as little as single copy but want that pg number…Is there enough material to do a test for presumptive blood? Maybe not.. Then do DNA test.When DNA extracted.. Want to release it from cell..Do laundry.. Take shirt with blood stain .. Want stain but not shirt..Keep dirty water..DNA free floating in it.. How it may be done

Do PCR in cycles38th cycle.. 536 million copiesTargeting STR’s 7 and 8 repeatsAmplify these targetsVariation.. PolymorphicReliable/reproducible

Inheritance of STR locus: paternal types 5,6 maternal types, 4,7 see in textbookChild can be any variety of combinations of these

Did OJ do it? RFLP results not usefulPCR yielded more samples First generation of testing faulty so certainty lacking All 13 STRs may not have been runCouldn’t justify that lab had doneThis data would give completely different results today

Defense team tore up DNA experts- this alone justified acquittal

Mixtures of DNA samples? How handled?New researchDNA, firearms, all trace evidence used to determinewho has best evidence.. If limitedLatent print analysis less successful than believedPresence of a fp..bare handsOils and aa , skin clings on printCan get DNA

Fire Debris AnalysisDespite a potential loss, some DNA can remain on explosives and fire startersEven if detonatedDNA more powerful evidence than any other

Drug chemistry and DNAIdentification of illegal drugsDrug e.g. cocaine- plant DNA source may be identifiedORDistributors touched the bag

Subway bombing in Madrid- read case- DNA did what ?

70% of people are shedders.. 30% are not

Mt DNA -Maternal -useful for degraded DNA samples Y chromosome STRS- Paternal inheritance Useful for mixtures or aspermatic samplesUsed to determine male lineagesLow template DNA- obtain profiles from samples with few cellsNYC good at this- why?Extremely controversial

Contamination susceptibleUsed on Lil’ Wayne gun case- ‘low copy’Take sample from gun.. Type many samplesOr one allele amplifies well.. Then do PCR

DNA from hair.. Without root tissue not as goodNon rooted hair.. Mt DNA shows maternal lineageJesse James and Oetzi the icemanSuper Bowl footballs- DNA testing certifies authenticityHuman genetic code of athletes- AustraliaAdded to ink used to mark all official goods- authenticityInclude in original artwork -artist DNASnowball the cat.. Mt DNADNA artCan get your profile on the wallResources:John Butler (not musician)Forensic DNA typing testsDNA criminalistics- Keith Inman and Norah RudinInman org site

Results analyzed thorough many trials and reviewsCost: PCR kit. $4k/ kit.. One sample $4k reagent control- $1000 Some evidence in case may have 100 or more samples - EXPENSIVE

Defense team lab work?Many times defense defers to DNA lab tests rarely disputed that DNA presentBut circumstances differe.g. Not rape- consensual.He was at party/cut himself cutting a lime.

PCR process targets only human DNA … that’s it how it can be distinguished from cat or dog or plant

How to determine if DNA is recent or old? Has been there on item?? Most DNA on item is probably last touched.. Or handled.

Samples preserved in perpetuity. Never destroyed….

Research Projects‘Outside the box’ of criminology addressed by DNA analysisBioforensics Solutions Lab: Geolocation.. Travel history of individuals, plant sources, genetic Identity. Individual and population specificAnd Exposure.. Environmental Conditions.. Experienced.. Epigenetics

As an organism grows and develops, carefully orchestrated chemical reactions activate and deactivate parts of the genome at strategic times and in specific locations. Epigenetics is the study of these chemical reactions and the factors that influence them. (Taken from Learn.Genetics)

Awesome..EPIGENETICALLY can distinguish between identical twins.ALL life ..

Wine/ grape genetics.. Powerful.. Agricultural applications.. Infectious diseases.. Etc.Thoroughbred horses, mosquito bites.. Carrier, our own DNA in insect populations from blood bites.Robotics enables science to handle DNA

Next Generation Sequencer

Generating 50 mega bases.. Takes no time now.. Would have taken 50 years..

Human genome examination. Very practical. Cheaper.High resolution melts.. Rapid.. Simple and low cost..rather capillary electro instrument.. Use PCR/HRM.. Expensive though.High Res Melt. Used to thermally separate double stranded DNA.. At hydrogen bonds..energy required to break bonds and sequence of bases will create a unique signatureMakes a melting profileNext generation seq data is informative and will be next processing in forensics

Has been decoded -computers have to handle data

Universal DNA typing scenarioStrange things in world..Lets say weapons cache found in PakistanBut this is not Pakistany in origin

Universal DNA typing could be a large projectAble to leverage DNA to determine origin and exchange of a particular cache of weapon

Leverage all generic informationHandling a package or anything.. Direct contact.. Leave DNAPollen, fp on tape.. Biological fluids on packaging.. Dog hair.. All exterior surfaces may be hotCan determine origin of item, travel routes, corroborating evidence.

Applied to some plants in Colombia- Coca plants… marijuana DNA of plants may be traces to fields and travel distribution.

Have material or person of concern Have particulates of concern pollen or plant. Remove DNA do PCR or HRM based plant ID. Then get computational information. Can see dispersal of these plants. Trafficking, routes, follow the DNA. Proof of Concept:Used shoes to test this project‘operation shoe phone’Took shoe at beginning of work day. Decontaminate with bleach and alcohol and walked around with shoe at work all day.Take sample of shoe and find unique signatures.They sample plants and animals to determine track.One person stayed in building but all others went out during day

Do not extend to prokaryotes. Too complexStuck to eukaryotes.. Specifically plants and animalsResults.. Found each of species present in each shoe. Number of species of plant and animals. Similar.42 % plants.. 58 % animal

59 million bases of sequences can use universal DNA profiling to distinguish geographical areas.

Physical GEOLOCATION SYSTEM: Bio information that are entrapped on person or thing as they travelRapid analysis of unique DNA fp of particulates

Spatial Decision Support SystemTravel routes based on public data, private routes and indicator speciesUSE specific local fauna or flora

Use Google Earth to map the routes.Amount of ecological data is in Google site(countries like Syria, China, Eastern Europe not as good data)

DNA mixture InterpretationEnhance ability to separate contributors to mixturesIncreased sensitivity of these kits. Helps to detect single cell scenarios .. SO sensitiveBUT with mixtures, comes difficult interpretationsCan apply to agricultureRisk to national security is food import and water… disease can be carried so easilycould wipe out crops quicklyDNA has a mixture when gathered in environment.What is the max number of alleles an individual can have.. 2 max One from each parent.Two copies could be same allele elementsPeak heights proportional to amount of DNA in sample.

May use machine learning or artificial intelligenceTrain a computer to index/ identify that we cannot .

How does it work? Have a question. Will it rain tomorrow? We ask ourselves a series of ?s. Is pressure dropping. Did it rain today or yesterday? Direction of wind? Etc..Computer program written to rate the ?s and evaluate this.. Indexing..

Threshold set by system to value each answer to ?s.. Decision is made.

All this to ask.. Who are the individual donors of the mixture of DNA?Programs try to capture human and computer interpretation

Touch DNA analysis

Factors involvedWhat is the item.. Sponge? Or glass item or handgun with friction ridges on grips? This mattersMatte or smooth surface?How is handled? How is item gripped or treated.? What are you doing with it?Shedder or non shedder is person?Duration of handling?(last to touch or one who touches most)Environment heat? Humidity sunlight ?Owner of itemDomestic or work or outdoor?The bottleneck is time and resources. Due to all factors hereNOT practical in labHave all items and kitchen sinkThis project tries to optimize sampling method..Used variety of ways to extract DNA .. None appropriate for all . So use different methods to cover possibilities

Touch DNA analysisIllegal materials cross into US.. Effort to prevent entering to countryForensic Analysis may be used to provide intelligence to deter entranceCustom handlers involved

Scientific Innovation: basic science, justice system, and national securityCan identify high impact and successful research Where interests intersect: Not just forensics and not just basic academic research