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Molecular Forensic

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11 views19 pages

Molecular Forensic

Uploaded by

tanishapatel1005
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Molecular: Forensics

Kyle Vinci P. Solano RMT, MD, DPSP


Advantages of DNA Testing in Forensics
• Direct Identification: DNA uniquely identifies individuals, using the 0.1% genome difference.
• Robustness: DNA is resilient to environmental factors; PCR enables results from trace amounts.
• Human-Specific: Testing targets human DNA, unaffected by bacterial contamination.
• Y-Chromosome & mtDNA: Useful for male-specific analysis and degraded samples, respectively.

DNA Sample Collection Techniques


Forensic Collection:
• Use sterile swabs for fluids, double-swab technique for bite marks and dried materials.
• Label and package each item separately to prevent contamination.

Sample Types:
• Buccal swabs, blood stains, or fresh blood for reference standards.
• In cases of decomposition, DNA from tissues like bone or teeth is preferred due to stability.
• Blood Collection:
• White blood cells provide DNA as mature red blood cells lack nuclear DNA.
DNA Preservation and Handling
Storage:
• DNA samples can be preserved at room temperature, 4°C, or -20°C depending on
sample type.
• Formalin-treated samples are suboptimal, but PCR can still yield results.

Contamination Prevention:
• Single-use gloves, clean instruments, and masks reduce contamination risk during
collection.
• DNA Degradation and Resilience

Factors in Degradation:
• DNA degrades via random fragmentation but remains analyzable for years if frozen
or dried.
Environmental Impact:
• Certain conditions, like metal ions and UV light, accelerate DNA degradation.

Legal Relevance
• DNA in Court: Forensic DNA results play a crucial role in legal cases; proper
specimen handling is critical for reliable evidence.
DNA Extraction and Quantification in Forensic Testing: Key Points
DNA Extraction
• Essential first step for DNA testing to isolate DNA from blood, biological stains, or other
biological sources.
• Various methods: solid-phase column extraction, magnetic resin particles, and silicon beads for
removing contaminants.
• Automated techniques commonly use column and magnetic particle methods for high-purity
DNA.
• Differential lysis separates male (sperm) DNA from female (epithelial) DNA, crucial for certain
samples like vaginal swabs.

Quantification
• Necessary for forensic STR testing to determine the appropriate amount of DNA for optimal
amplification.
• Real-time quantitative PCR offers both qualitative and quantitative assessments of total human
and male DNA.
• Insufficient samples may halt testing at the quantification step, especially in cases with limited
Genetic Marker Systems and Testing Methods
Marker Requirements
• Genetic markers for testing need high exclusion power, stable expression, and known
mutation rates.
• Codominant expression, minimal null alleles, and stable storage conditions are key for
reliability.
• Genetic markers should have established gene frequencies across populations for accurate
paternity/identity calculations.

DNA Polymorphisms
• Variations between individuals are due to DNA polymorphisms; about 1 in 1300 bases differ.
• Polymorphisms in noncoding "junk" DNA, especially in repetitive regions, are critical for
forensic analysis.
• Variable Number of Tandem Repeats (VNTRs) and Short Tandem Repeats (STRs) are used for
length-based polymorphisms.
• Sequence polymorphisms include SNPs and sequence-based tests, enabling detailed forensic
analysis.
Short Tandem Repeats (STRs)
STR Loci
• STR loci contain short repeat sequences that vary among individuals, commonly used in forensic
DNA profiling.
• Amplification involves PCR with labeled primers, allowing detection through capillary
electrophoresis.
• The 20 core STR loci in CODIS create genetic profiles with high accuracy and a low probability of
random match.
• STR analysis is highly discriminative, with random match probabilities often rarer than 1 in 1
trillion among unrelated individuals.

X and Y Chromosome Markers


Sex Determination and Y-STRs
• Amelogenin locus identifies sex by amplifying specific bands on X and Y chromosomes.
• Y-STR markers are useful in mixed male-female samples, enabling clear male profiles without
female interference.
• Y-STRs are also used for paternal lineage tracing and can identify human remains through
Mitochondrial DNA Sequencing
• Used for extremely small or degraded DNA samples (e.g., shed hairs, skeletal remains).
• Hundreds to thousands of copies of mtDNA per cell; only one copy of nuclear DNA.
• Can be useful when nuclear DNA is unavailable.
• mtDNA is maternally inherited and has no paternal contribution, providing a unique
match for maternal lineage.
• Analysis focuses on the noncoding displacement loop (D-loop), with hypervariable
regions.

Single Nucleotide Polymorphisms (SNPs) and Next-Generation Sequencing (NGS)


• SNPs detect base changes in DNA and can provide information on relatedness, traits, and
ethnic origin.
• NGS allows for high-throughput sequencing, offering a faster, more cost-effective method
than traditional sequencing.
• NGS can detect SNPs and STRs simultaneously, providing more precise results than size-
based capillary electrophoresis.
Use of DNA Evidence in the Crime Laboratory
• DNA tests can exonerate wrongfully accused suspects or link suspects to crimes.
• Common applications include sexual assault, homicide investigations, and accident
reconstructions.
• DNA can help identify human remains, link serial crimes, and solve cold cases.

Familial and Genealogy Testing


• Familial testing used to identify relatives of perpetrators, with Y-STRs being particularly
useful for male lineages.
• The Grim Sleeper case (2010) identified a serial killer through familial DNA testing.
• Forensic genealogy gained public attention in 2018 after the Golden State Killer was
identified using genealogical databases.
• Consumer DNA databases (e.g., Ancestry.com) are being used to match crime scene
DNA with potential relatives, leading to breakthroughs in cold cases.
• Ethical and privacy concerns exist regarding the use of public DNA databases in criminal
Future DNA Testing (Rapid DNA Testing):
• Goal: Automate DNA profiling at booking stations for quick DNA analysis within hours.
• DNA profile uploaded to NDIS and searched against unsolved crimes while the arrestee is in custody.

DNA in Pathology Laboratories:


• Used to resolve specimen mixups (e.g., switch or contamination of biopsy samples).
• Can identify "floater" cancerous tissue on microscope slides, verifying origin.
• Used in challenging urine drug tests or verifying sample authenticity.
• STR markers help evaluate marrow engraftment post-transplant.

Exclusion of Parentage:
• DNA tests used to identify biological parents, with >99% accuracy.

Exclusion types:
• Direct Exclusion: Identifies nonparentage when a child’s allele isn't present in the presumed parent.
• Indirect Exclusion (Reverse Homozygosity): When an expected genetic marker is absent in the parent,
indicating nonpaternity but not conclusive.
Probability of Exclusion (PE):
• PE estimates the chance of excluding a random individual from parentage based
on genetic alleles.
• Higher PE in more polymorphic genetic systems.
• Combined Probability of Exclusion (CPE): Calculated using multiple genetic
systems, yielding a CPE >0.995.

Inclusion of Parentage:
• If no exclusion occurs across multiple tests, a paternity estimate is calculated.
• The tested male may still be falsely accused; thus, further testing is necessary.
• Calculations depend on gene frequencies, and various populations' frequencies
may affect results.
Paternity Index (PI) Calculation:
• Likelihood ratios (x/y) compare the tested male to the random population for paternity
estimation.
• PI >100 indicates strong evidence of paternity (99:1 odds).
• PI values have legal significance and may shift the burden of disproving paternity.

Parentage with an Absent Parent:


• If one parent is unavailable, calculations for parentage can still be done based on the alleles
shared by the child and the tested male.
• Complicated by racial differences between the missing parent and tested individual.

Reconstruction of Families:
• DNA tests help establish familial relationships in cases of disputed inheritance or
identification.
• Important in mass disasters, establishing parental relationships, or immigration status.
• Reconstructing family relations can be done using multiple relatives or testing biological
fin

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