Mid Term 2
Mid Term 2 100
Popular in Forensic Science 100
Popular in Criminology and Criminal Justice
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Frnscmo M T2 Serology Crimesevidence amp Forensic Serology Violent Crimes Murder Rape Assault White collar crimes Mail fraud Bank robbery Forensic serology is the detection classification and study of various bodily fluids such as blood semen amp saliva and their relationship to a crime scene Serology is the first step in stain identification Source attribution of stains will be done by DNA testing following serological examination Presumptive tests sensitive but not specific will react with other compounds Confirmatory tests specific but less sensitive eg Blood Identification Presumptive Phenolphthalein luminol Confirmatory Takayama Test Takayama is a crystal test used for the confirmation of the presence of blood on samples that screened positive a presumptive test It reacts with Hemoglobin in blood Semen Identification Presumptive Acid Phosphatase Confirmatory Sperm P30 Saliva Identification Presumptive Amylase Blood Blood is a highly complex mixture of cells enzymes proteins and inorganic substances Plasma the fluid portion of blood composed principally of water Red blood cells erythrocytes Antigens usually proteins are located on the surface of red blood cells and are responsible for bloodtype characteristics White blood cells leukocytes Contains DNA Platelets are the solid materials suspended in plasma Blood Purpose 0 Supplies oxygen to tissues bound to hemoglobin which is carried in red cells 0 Supplies nutrients such as glucose amino acids and fatty acids dissolved in the blood or bound to plasma proteins eg blood lipids o Removes waste such as carbon dioxide urea and lactic acid 0 Responsible for immunological functions including circulation of white blood cells and detection of foreign material by antibodies 0 Coagulation part of the body39s selfrepair mechanism blood clotting after an open wound in order to stop bleeding 0 Transport of Hormones 0 Regulation of body pH 0 Regulation of core body temperature Blood Identification Phenolphthalein Test Presumptive test Chemical tests which uses colour change to detect the possible presence of blood Phenolphthalein KastIeMeyer test relies on the iron in hemoglobin which is the ironcontaining portion of a red blood cell to promote the oxidation of phenolphthalin to phenolphthalein Phenolphthalin is colorless but in the presence of blood and hydrogen peroxide it changes to phenolphthalein which makes the solution pink This test is very sensitive One drop of blood diluted in 10000 drops of water can still be detected by the KastIeMeyer test will give falsepositive result in the presence of vegetable peroxidases such as those in horseradish broccoli cauliflower etc other oxidizing species in the sample will also cause a false positive chemical oxidants such as copper and nickel salts will cause the KastleMeyer reagent to turn pink before the addition of the hydrogen peroxide thus it is vitally important to add the reagent first then wait a few seconds then add the hydrogen peroxide Luminol Test Luminol C8H7N302 is a chemical that exhibits chemiluminescence with a striking blue glow when mixed with an appropriate oxidizing agent like heme in blood thus it may be used to locate traces of blood even if it has been cleaned or removed The solution of luminol and the activator are sprayed on the area under investigation The iron present in any blood in the area catalyzes the chemical reaction that leads to the luminescence revealing the location of the blood The amount of blood necessary for the reaction to occur is very small relative to the amount of luminol allowing the detection of even trace amounts of blood The glow lasts for about 30 seconds and is blue Detecting the glow requires a fairly dark room Any glow detected may be documented by a long exposure photograph The room must be very dark in order to visualize the luminescence Photography of the reaction is critical and can be very difficult The luminal also is sprayed and can be difficult to perform in non ventilated spaces Luminol Test This is a presumptive test It will give a false positive results and it can be triggered by a number of substances such as copper or copper containing alloys certain bleaches and as a result if a crime scene is thoroughly cleaned with a bleach solution residual cleaner will cause the entire crime scene to produce the typical blue glow effectively camouflaging any organic evidence such as blood small amounts of blood present in urine and it can be distorted if animal blood is present in the room that is being tested fecal matter causing the same glow as if it were blood It may prevent other tests from being performed on a piece of evidence Ludwik Karol Teichmann 18231895 Polish physicianchemist In 1853 published a paper on the crystallization of certain organic compounds of the blood describing the preparation of the microscopic crystals of hemin heme chloride First test for the presence of blood in suspect stains on clothes furniture or other objects for blood the suspected liquid is put under a coverglass with a crystal of sodium chloride and a little glacial acetic acid heat carefully without boiling and then cool If blood is present rhombic crystals of hemin will appeah Karl Landsteiner Vienna physician and professor 19011909 working with blood transfusions that some individuals blood clumped discovered that the blood can be typed into different groups These groups are eventually labeled as types A B AB and 0 first suggested these could be used to establish paternity antibodies and antigens introduce chemistry into serology 1930 awarded the Nobel Prize ABC Typing System A blood type also called a blood group is classification of blood based on the presence or absence inherited substances on the surface of red blood cells RBCs and the presence of antibodies in the plasma RBCs may have A antigen B antigen both A amp B or neither A amp B antigen The presence or absence will determine a person39s blood type Type A will have A antigens on RBC and anti B antibodies in serum Type B will have B antigens on RBC and anti A antibodies in serum Type AB will have A amp B antigens on RBC and no antibodies in serum Type 0 will have no antigens on RBC and A amp B antibodies in serum People with Type 0 blood are considered the universal donors because their blood can be transfused into anyone without causing reactions Why was it used ABO typing can be used to sort people into four groups Reference blood samples are typed and compared to unknown blood stains Type is different reference is excluded as a possible source of the stain Type is the same reference is a possible source of the stain but not conclusive because other people also have the same type Statistics needed to be reported with the result Leon Lattes o Lattes thought that ABO blood typing might be useful for the identification of individuals 0 Professor at the Institute of Forensic Medicine at the University of Turein in Italy 0 1915 he developed procedure for determining the blood type ABC of a dried blood stain 0 Applied it to criminal investigations Types of Sexual Assault Evidence Swabs Vaginal anal oral surface and penile Generally have multiple sources of DNA Slides confirm presence of sperm Public Combings roots vs shafts microscopy DNA analysis Articles of Clothing bedsheets undergarments towelwash clothes alternate light sources for finding stains Semen Semen is an organic fluid also known as seminal fluid that may contain spermatozoa sperm Finding the presence of sperm is a confirmatory test for semen In humans seminal fluid contains several components besides sperm proteolytic and other enzymes such as acid phosphatase AP The level of AP activity is 500 to 1000 times higher in human semen than in any other normal body fluids or secretions The presence AP is sometimes used for presumptive testing for semen Prostate Specific Antigen PSA also known as p30is present in small quantities in the serum of men with healthy prostates but is often elevated in the presence of prostate cancer and in other prostate disorders The presence of PSA p30 is used for confirmatory testing for semen Sperm Head Contains NucDNA MidSectionNeck Contains mtDNA which supplies energy to cell Tail Protein bers that contract and release on alt sides to create the swimming motion lOOO complete movements to swim 125cm In uences on Finding Sperm Vasectomy or low sperm count Minimal ejaculation Urination Vaginal pH Contraceptive usage Bathing Survival of Sperm Important that samples are collected as soon as possible as sperm survive for different periods of time depending on the environment they are found in 72 hours to 7 days in the vaginal cavity 24 hours to 3 days in the anal cavity 824 hours in the oral cavity Semen Identification To make searching for sperm easier prep for microscopy Extraction buffer plus SDS and Pro K may be added to a sperm sample to lyse any nonsperm cells epithelial white blood Sperm heads are largely unaffected Sperm tails can be quotlysedquot or stripped from the sperm cell H amp E Staining Hematoxylin and Eosin Hematoxylin a basic dye binds to acidic components of a tissue such as DNA and RNA usually a blue or purple color Eosin an acidic dye binds to basic components of a tissue usually a red or pink color 1Simple test Extract stain into a buffer Place drop on test strip in quotSquot well Negative no color Positive pink color Semen Identification 2 ABAeards Detects the presence of p30 protein in seminal fluid named because it weighs 30000 daltons p30 is 200000 to 55 million nanogram per ml of semen The sensitivity of ABAcard p30 test is 4 ngml p30 reacts with mobile monoclonal antihuman and forms mobile AbAg complex AbAg migrates to test area T and binds with immobilized antibody anti p30 When AbAg Ab gt 4 ngml pink dye particles colored band in the test area T If no p30 is present mobile Ab migrates to C area is imobilized and forms a pink band to show test is working Saliva Saliva is rich in the enzyme alphaamylase aka ocamylase salivary amylase an enzyme that breaks down complex carbohydrates into smaller sugar molecules The presence of amylase is presumptive for saliva as it is also present in fecal material and vaginal secretions Amylase is an enzyme and may become inactive This would give a false negative reaction for the presence of saliva Presumptive tests include 1 StarchIodine test for amylase Amylase will breakdown starch Iodine is added and will detect starch that has not been hydrolyzed Amylase radial diffusion assay Pour agar gel containing starch into culture plate Create wells in solidified agar with vacuum punch Add extract of evidence sample to well Add standards to other wes amylase samples at known concentrations Incubate overnight at 37 degrees C Stain with iodine solution Not specific test for amylase 2 Phadebas 3 Rapid Stain Identification RSID Saliva Saliva Presumptive Tests StarchIodine test for amylase Amylase will breakdown starch Iodine is added and will detect starch that has not been hydrolyzed Amylase radial diffusion assay Pour agar gel containing starch into culture plate Create wells in solidified agar with vacuum punch Add extract of evidence sample to well Add standards to other wells amylase samples at known concentrations Incubate overnight at 37 degrees C Stain with iodine solution Not specific test for amylase 2 Phadebas Phadebas tablets consist of insoluble starch polymers to which a blue dye has been covalently bound Extract sample and add tablet When amylase is present Polymers are degraded Dye is liberated and becomes soluble Concentration of soluble dye is measured by spectrophotometry at 620 nm 3 Rapid Stain Identification RSID Saliva One such test is the RSIDSaliva kit which is a lateral flow chromatographic strip test designed to detect a amylase an enzyme present in human saliva Designed to work like the p30 test DNA Nuclear Deoxyribonucleic Acid DNA A living things have DNA which is found in the cells of those living things DNA acts as a blue print or instruction manual for the living organism Forensic DNA typing involves understanding 1 Structure of DNA 2 Techniques involved in DNA testing 3 Uniqueness of DNA DNA found in the nucleus of the cell is called nuclear DNA DNA found in mitochondria of the cell is called mitochondrial DNA DNA is Inherited A child39s nuclear DNA is inherited from both parents A child will receive half of their nuclear DNA from their Mother and half of their nuclear DNA will come from their father Chromosomes The DNA is organized in the cell into packets of information known as chromosomes Humans have 23 pairs of chromosomes We inherit half of our chromosome pairs from each parent The 23rd pair are the sex chromosomes X and Y These determine the gender of the person Females have two X chromosomes and males have an X and a Y chromosome DNA Nuclear Genes Regions of DNA that do something or code for something are known as genes Thus DNA is often called genetic material Genes are very important regions of the DNA because if there are any changes in these regions then often the person will suffer some type of disease Scientists who study genetic diseases focus on trying to determine what areas of the DNA are gene regions and what changes may happen to these area that will cause disease Forensic scientists have been able to use the same techniques used by these genetic scientists to develop forensic tests for DNA typing Reasons for DNA testing Establish paternity andor maternity Use known samples from close relatives to help determine DNA profiles of a missing person or a crime victim Twins Identical Twins will have the exact same DNA type Fraternal Twins will not because two eggs are fertilized Everyone else will have a unique DNA pattern DNA Structure Deoxyribonucleic Acid DNA is a double helix strand that is composed of Deoxyribose sugar group rails of the ladder Phosphate rails of the ladder Nitrogenous bases rungs of the ladder present in DNA 1 Adenine A 2 Thymine T Pu rines 3 Guanine G 4 CtosineC 5 Ullacip lerimidines found in RNA not DNA Bases described differently depending on What molecules are attached to them Nucleotides if sugar and phosphate groups are attached Adenine Adenylate Guanine Gyanylate CytosineCytidylate ThymineThymidylate Why test DNA from evidence It can be retrieved from evidentiary stains blood semen perspiration and tissues DNA is very stable DNA tests and procedures that have been used to conduct genetic research are available for the forensic scientist to use and apply DNA is Unique unless the source has an identical twin and can be used to determine the source of the stain if sufficient regionsareas of the DNA are examined Potential DNA Evidence Body Fluids cells blood semen saliva perspiration Tissue bones teeth organs Waste Excretions urine fecal material Dried Blood stains dried semen stains vaginal swabs undenNear clothing sheets dried saliva cigarette butts stamps envelope flaps F erNT DNA Profiling is a Comparison DNA Complexity DNA Typing Compare DNA types in evidence with DNA types in from reference samples from suspects or victims Reference samples may be a blood sample Reference samples may be a saliva sample If the types match the source of the reference sample is a possible source of the DNA in the evidence If enough DNA is tested then the source of the stain will be identified If the types do not match the source of the reference sample is excluded Complexity Each human cell contains over six feet of DNA It is a very large molecule with approximately 3 billion base pairs Approximately 23000 human genes Over 98 of human DNA is the same across individuals Forensic DNA testing is directed at regions of DNA where there are differences in the DNA Typically those test will avoid gene regions because there will not be a lot of differences in these areas of the DNA DNA Typing DNA typing procedures exploit areasregions of DNA that are polymorphic eg HumanEye colour hair colour blood type DNA Polymorphisms Sequence and length These regions are areas where there will be differences betweenamong individuals DNA typing looks at different loci and figures out which alleles you have gotten This is your DNA type Current PCR based DNA typing systems will look at 1316 different loci Locus Loci amp Alleles Locationarea on a particular chromosome is called a An allele is the version of DNA that you find at a particular locus The plural is loci locus Sometimes this version of DNA will confer a trait like blue or The locus IS named according to Its posmon on a brown eyes Sometimes it does not chromosome Some loci are named because of a disease that occurs due to geneth difference as that You will get one allele from your mom and one allele from particular location on the chromosome your dad If they are the same you are homozygous Forensic DNA typing looks at loci where there is If they are d39fferent you are heterozygous polymorphism Variable Number Tandem Repeats VNTRS The VNTR39s may have 245 bases in the repeating sequence The more repeats there are the longer the piece of DNA will be This works for both RFLP and PCR based STR typing The example above shows that the 5 allele is shorter than the 7 allele Alec J effreys 1950 English Genetics professor 19834Believed his DNA analytical technique can be used to individualize a person Works with local police to obtain DNA from crime scene stains and compares to blood samples given by all males in a village This work revolutionizes the use of DNA in forensic serology Was knighted for his work Topic of the book The Blooding Restriction Fragment Length Polymorphism RFLP Testing 1 Oldest test First used in 1988 2 Very slow process weeks or days 3 Need good quality DNA 4 Need large amount of DNA 5 Separated DNA based on length polymorphism Steps Extraction Remove DNA from inside the cell Process DNA Cut the DNA with restriction enzymes Separat e the fragments of DNA according to size electrophoresis Visualize what has been separated using specially designed DNA probes that are specific to the loci of interest Issues RFLP testing required large stains about the size of a nickle that were relatively fresh Usually only successful with rape cases and homicides with substantial stains Did not work on old stains because the DNA would often be too degraded Took weeks or days to complete the tests It was very informative testing if there was enough good quality DNA Polymerase Chain Reaction PCR PCR techniques dramatically advanced forensic DNA typing capabm es 1 All current DNA methods are PCR based 2 Works on degraded DNA evidence 3 Works on small amounts of DNA 4 Current methods examine length polymorphisms The developer of PCR technology Dr Kary Mullis was awarded a Nobel Prize for his work in 1993 Evidence suitable for PCR Typing PCR tested allowed for testing of very small amounts of DNA and degraded DNA 80 small blood stains saliva perspiration stains would have enough DNA to test with PCR cigarette butts stamps envelops chewing gum threads baseball caps ski masks headbands small blood spatter fingernail clippings toothbrushes hair brushes eyeglasses phone receivers pens false teeth Polymerase Chain Reaction PCR How it works DNA is extracted from evidentiary stains Extracted DNA amplified using PCR Amp1ified regions of DNA are typed according to size PCR Ingredients Extracted DNA Primers ATCGs Taq enzyme Buffer PCR is a series of reactions that copies areas of the DNA molecule These areas are preselected based on the purpose of the test PCR amplification is a process used by molecular biologists for research There are three steps in PCR 1 Denaturation DNA molecule is heated and the DNA ladder splits in half 2 Annealing Special small pieces of DNA called primers that are designed using complimentary base pairs find the region of DNA that is to be copies The DNA that has been split in half is cooled slightly so that the primers will bind with the DNA 3 Extension The DNA strand that has been split apart will now be copied using a special enzyme called Taq polymerase Taq will read the strand and add dNTPs the building blocks of DNA PCR Equipment Thermal Cycler The thermal cycler is a key piece of equipment It is a very special heat block that can raise and lower the temperature very accurately The tubes are placed in the thermal cycler which will heat and cool in cycles so that the PCR reaction can occur in each of the tubes Step 1 DENATURE The Tubes are heated and the DNA molecule wi unzip into two halves Step 2 ANNEAL The tubes are cooled slightly and the primers that are in the tubes with the unzipped DNA will bind to the complementary site Primers are small DNA pieces that locate the locus of interest that will be copied Step 3 ELONGATEthe unzipped strand will be copied because the Taq polymerase enzyme will read the strand of DNA and add the appropriate dNTP that matches The temperature is just a little bit hotter than the primer stage but not too hot or the dNTPs won39t bind Replication copying happens through a process called Thermal Cycling because the heat level changes A special piece of equipment called a Thermal Cycler is used to carry out this process Forensic PCR Tests The first PCR based typing tests were based on sequence differences dots DQ alpha typing Polymarker The current PCR based typing tests are based on length differences STR s Capillary Electrophoresis used to separate amplified DNA fragments according to their size Statistics of DNA Typing The population studies determine how many times the alleles will be found in a particular population Those allele frequencies can be mathematically combined to determine the rarity of the overall DNA type at each locus and then multiplied across all loci to give the overall frequency of the DNA genotype DNA Mitochondrial Y STRS Mitochondria organelle found in most eukaryotic cells large number in certain cells 1000s quotcellular power plantsquot generate most of the cell39s supply of ATP implicated in several human diseases involved in aging Critical to cell function Have their own blueprint Mitochondrial DNA mtDNA mtDNA is located in each mitochondria 2 to 20 copies are in each Smaller than nuclear DNA only 16569 base pairs in size it is circular DNA still has the double helix structure of DNA justjoined in a circle first significant part of the human genome to be sequenced mtDNA is maternally inherited because mtDNA is passed from mothers to their children The female children will pass the same information to their children same for all maternal relatives Circular Chromosome Forensic mtDNA typing concentrates on the control region where there is some variability Hypervariable region I Hypervariable region ll These are the regions typed mtDNA Structure Even though mtDNA is circular the basic structure is still the same and so PCR amplification can still be conducted on mtDNA to amplify the regions of interest HVI HVII mtDNA and forensic DNA Typing mtDNA typing is used on very degraded samples or samples with very little nuclear DNA hair bone and teeth becauseit Works with a limited sample Due to high copy number more copies of mitochondria in one cell and more copies of mtDNA in each mitochondria Works with degraded samples Less prone to degradation than nuclear DNA structure location Maternal inheritance maternal relatives can be the source of a reference sample in missing persons cases Some variability between individuals who are not maternally related allows for differentiation among samples Disadvantages Is not a unique DNA type because all maternal relatives will have the same mtDNA type It is not as discriminating as nuclear DNA type Typing technique requires DNA sequencing which is more complicated that current nuclear typing procedures mtDNA typing involves amplification of DNA and it is very prone to contamination so DNA extraction is done in a very clean and controlled environment The hood on the bench is used to isolate the samples Before extraction of the DNA the hair needs to be processed using small tissue grinders that are specially designed for this process Sequencing The sequence of the bases in the HV l and HV ll regions are determined by analysis They are compared to a reference sequence that was first determined by Anderson It is now known as the Cambridge Reference Sequence CRS The differences between the aligned samples and the CRS are the only things noted all other positions in the sequence are understood to be the same as the CRS For example position number 16311 in the CRS is listed as a thymidine or T However in some individuals that position could be substituted with a cytosine or C This polymorphism is designated as 16311 C and all other positions in the region sequenced are understood to be identical to the CRS Compared to reference samples The sequences are compared to each other and to any reference sequences to see if there are any matches The interpretation of a match is affected by a biological event called heteroplasmy that can cause samples from the same person to have slight sequence differences in a position Heteroplasmy detecting more than one mtDNA type within an individual There are so many copies of mtDNA in the various tissues that occasionally there will be a one base pair difference at a position in a tissue blood sample or between tissues hair and blood Armed Forces DNA Identification Laboratory AFDIL The Armed Forces DNA Identification Laboratory AFDIL is a forensics laboratory specializing in DNA profiling run by the United States Armed Forces AFDIL stores refrigerated DNA samples from all current active duty and reserve personnel AFDIL will use DNA testing to identify remains of soldiers killed in action Provides worldwide scientific consultation research and education services in the field of forensic DNA analysis to the Department of Defense and other agencies Current mtDNA protocols were developed at AFDIL by Dr Holland and others Lineage Markers Lineage markers are useful for assisting with the identification of related individuals separated by many generational events Applications of YSTR Testing In Forensrcs Historical cases identification Sexual assault evidence when conventional STR analysis fails or is problematic typically up to 72 hrs post coitus Verification of gender if amelogenin results are unclear Missing persons investigation assist in identification of human remains Paternity testing motherless cases Haplotype A haplotype is a series of single alleles from different loci It is a single allele at each locus because it is a haploid system not a diploid system like with a nuclear type where one allele comes from Mom and one comes from Dad There will only be one allele for each locus that is typed YSTR profiles and mtDNA sequences are considered haplotypes Frequency of YSTR haplotype is determined from a database just as with mtDNA Y STR Typing amp Y Chromosome Typing All males will have the same Ystr type because females do not have a Y chromosome Can be used to further characterize male DNA Can be used to type mixtures of female and male DNA as happens in some rape cases Definitive Results Y STR exclusions are definitive proof that two individuals are NOT paternally related as are mtDNA exclusions for maternal relationships Not BLOOD relatives Limitations Y STR results CANNOT differentiate between paternally related individuals with the rare exception of mutational events PCR amplification of several STR loci on the Y chromosome will give a haplotype for the individual This typing is exactly like the typing process used to type nuclear DNA The primers are different and target DNA on Ychromosome Thomas Jefferson Case DNA Databases Justification In the early 90s the DNA community realized that DNA types could be used to do more than just confirm an individual was present or involved in a criminal act DNA types RFLP profiles stored in a database could be used to proactively link and solve crimes just like fingerprints were being used Also DNA could be used to identify human remains of missing persons Rapes could be linked together by DNA typing the various unknown semen from rapists and crime scenes This created investigative leads Statistics showed that rapists were quotrecidivistsquot ie they repeated their crimes overagain If a person was convicted of rape then their DNA could be stored in a database to be compared to quotunknown suspect rapes If they continued to rape women after their release from prison they would be caught again because their DNA type was in the database and cases would be solved Opposition The concept of DNA databasing was hotly contested and there was a strong opposition from the ACLU and attorneys such as Peter Neufeld and Berry Scheck These are the two attorneys who started project innocence Also involved in the OJ Simpson case Privacy concerns were at the center of the discussion Combined DNA Index System CODIS Began in 1990 as a collaboration among 14 forensic laboratories The FBI developed the CODIS software and then provided it for free to laboratories as they developed the capability to do DNA typing Each state had to pass DNA legislation in order to set up the DNA databasing capability It took several years for this to happen The DNA Identification Act of 1994 authorized the use of DNA data for forensic analysis and formalized CODIS It is the first Federal legislation concerning forensic Science By October 1998 CODIS became operational on a national level NDIS because all 50 states had passed laws allowing them to collect and store DNA profiles from convicted offenders As of 2004 all 50 states along with Puerto Rico the US Army and the FBI were CODIS participants States began expanding their DNA laws to include more categories of convicted offenders The original laws only collected from individuals who committed serious felonies like murder and rape For example the addition of DNA profiles from individuals arrested for burglary increased the number of cases linked together by DNA typing Today over 180 public law enforcement laboratories participate in NDIS across the United States Internationally more than 60 law enforcement laboratories in over 30 countries use the CODIS software for their own database initiatives International laboratories using the CODIS software do not have any connectivity to the United States CODIS system US Combined DNA Index System CODIS FBI sponsored software development Software provided free to all states and various countries States passed laws to establish their local and state databases Combined DNA Index System CODIS Addition of Arrestee s DNA into CODIS Some states have enhanced their DNA database laws to allow for the collection and comparison of DNA from people who are arrested for crimes This means that even though a person has not been convicted of a crime their DNA can be compared to profiles in the Forensic index If an arrestee39s profile matches they can then be charged with that crime There is currently not a federal law that allow for DNA collection from people arrested for a federal violation CODIS Hierarchy All three levels contain forensic and convicted offender indexes Local DNA Index System LDIS is installed at crime laboratories is operated by police departments or sheriffs39 offices DNA profiles originated at the local level can be transmitted to the State and national levels State DNA Index System SDIS allows local laboratories within that State to compare DNA profiles SDIS is operated by the agency responsible for implementing and monitoring compliance with the State39s convicted offender statute National DNA Index System NDIS is the highest level of CODIS enables qualified State laboratories that are actively participating in CODIS to compare DNA profiles NDIS is maintained by the FBI under the authority of the DNA Identification Act of 1994 CODIS Several Indexes Categorize the profiles Several Indexes Categorize the Profiles Entered into CODIS Convicted Offender contains DNA profiles of individuals convicted of crimes Arrestees contains profiles of arrested persons if state law permits the collection of arrestee samples Forensic contains DNA profiles developed from crime scene evidence such as semen stains or blood Missing Persons contains DNA reference profiles from missing persons In 2000 the FBI Laboratory developed the National Missing Person DNA Database NMPDD Program for the identification of missing and unidentified persons STR YSTR and mtDNA can be entered into the Missing Persons indexes of CODIS NMPDD uses 3 indexes in NDIS to enter DNA profiles that can be searched against each other Unidentified Human Remains Missing Persons Biological Relatives of Missing Persons Biological Relatives of Missing Persons contains DNA profiles voluntarily contributed from relatives of missing persons Unidentified Humans Remains contains DNA profiles developed from unidentified humans and remains CODIS Searches DNA profiles from an unknown sample from a crime can be searched against other unknown samples in the Forensic Index These searches link crimes together DNA profiles from an unknown sample from a crime can also be search within the Convicted Offender andor Arrestee Indices if that state is authorized to collect and database DNA samples from arrestees If there is a candidate match the laboratory will go through procedures to confirm the match and if confirmed will obtain the identity of the suspected perpetrator Once a match is identified the laboratories involved in the match exchange information to verify the match and establish coordination between their two agencies The match of the forensic sample against a record in the index may be used to establish probable cause to obtain an evidentiary DNA sample from the suspect The laboratory can then perform a DNA analysis on the known biological sample from the suspect so that this analysis can be presented as evidence in court Audit Document The FBI Laboratory developed a single comprehensive audit document Specific for monitoring compliance to the FBI Director39s quality assurance standards Applies to both casework and convicted offender DNA laboratories NDIS National Database Has STR nuclear and Y and mtDNA profiles in database contains over 10484400 offender profiles and 412500 forensic profiles as of January 2012 quotInvestigation Aided tracks the number of criminal investigations where CODIS has added value to the investigative process As ofJanuary 2012 CODIS has produced over 171800 hits assisting in more than 165100 investigations DNA Database accepted at NDIS NA data generated through PCR STR technology Y chromosome STR Y STR technology and mtDNA technology are accepted at NDIS Y STR and mtDNA data is only searched with the missing person related indexes Data Acceptance Rules The DNA data must be generated in accordance with the FBI Director s Quality Assurance Standards The DNA data must be generated by a laboratory that is accredited by an approved accrediting agency The DNA data must be generated by a laboratory that undergoes an external audit every two years to demonstrate compliance with the FBI Director s Quality Assurance Standards The DNA data must be one of the categories of data acceptable at NDIS such as convicted offender arrestee detainee legal forensic casework unidentified human remains missing person or a relative of missing person The DNA data must meet minimum loci requirements for the specimen category The DNA PCR data must be generated using PCR accepted kits and Participating laboratories must have and follow expungement procedures in accordance with federal law DNA Identification Act 1994 Established regulations controlling DNA forensicdatabase laboratories Provided for funding for establishmentimprovement of state and local forensic DNA Labs Required the FBI Director determine DNA standards for the community via DNA Advisory Board Standards issued in October of 1998 Compliance required for participation in NDIS and receipt of federal DNA funding All forensic DNA laboratories must be accredited within 2 years of issuance of the standards Participation in the National DNA Index and or receipt of federal funds for improvement required Compliance with the FBI Director s standards Protection of the privacy of the individual s DNA profiles in the database Mandatory external proficiency tests every 180 days FBI Director s DNA Standards Issued in October of 1998 Compliance required for participation in NDIS and receipt of federal DNA funding All forensic DNA laboratories must be accredited within 2 years of issuance of the standards FBI Director s Quality Standards Quality Assurance Program Organization and management Personnel Facilities Evidence Control Validation Analytical Procedures Equipment Calibration amp Maintenance Reports Proficiency Testing Corrective Action Audits Safety Subcontractors First Technical Working Group in Forensic Science TWGDAM Technical Working Group on DNA Analysis Methods TWG DAM Members of TWGDAM included forensic scientists from various local state and federal laboratories Issued QAQC Guidelinesin1991 These guidelines were very important in admissibility hearings concerning the admissibility of DNA typing Revised TWGDAM Guidelines were issued in 1995 to include mitochondrial DNA testing Became SWGDAM Scientific Working Groups on DNA Methods SWGDAM Formerly TWG DAM SWGDAM serves as a forum to discuss share and evaluate forensic biology methods protocols training and research to enhance forensic biology services as well as provide recommendations to the FBI Director on quality assurance standards for forensic DNA analysis Maintains and updates the FBI Director39s Quality Assurance Standards Maintains and updates the audit documents associated with the Quality Assurance Standards Familial Searches of DNA Databases DNA is inherited from parents so DNA profiles from siblings and off spring will share portions of a DNA profile quotFamilial Searching is an intentional or deliberate search of the database conducted after a routine search for the purpose of potentially identifying close biological relatives of the unknown forensic sample associated with the crime scene profile Familial Searching is still not done in every state as it has been controversial Three States conduct familial searching ColoradoDenver California Virginia Pennsylvania has legislation pending to consider familial searching eg Grim Sleeper Case in California solved Microbial Forensics Microbiology is the study of microscopic organisms or any living organism that is either a single cell unicellular a cell cluster or has no cells at all acellular Many microorganisms microbes are pathogenic and cause disease while other microbes are not a danger to human health Microbiologists use many tools to study microbes Microbiological techniques can be used to isolate and identify organisms of interest A common method is culturing the microbes by growing them in plates called petri plates using selective growth media The microbes will grow in colonies Media contains all the nutrients needed for the organisms to grow In order to identify the organisms they may be grown under special conditions such as with oxygen aerobic without oxygen anaerobic with antibiotics in growth media Observation of appearance of colonies means the microbe grows in those conditions Microbes are also observed microscopically using both light microscopes and electron microscopes Staining The Gram Stain Staining microbes with different chemicals allows the microbiologist to identify the organism One common stain used is called the Gram Stain Gram staining is used to differentiate bacterial species into two large groups Gram positive and Gram negative based on the physical properties of their cell walls Detects peptidoglycan present in a thick layer in Gram positive bacteria Microbial Forensics Microbial Forensics is an emerging interdisciplinary field of microbiology devoted to the development evaluation validation and application of methods to detect and fully characterize microbial samples containing a biological agent or its components Ability to identify viable pathogens Ability to identify DNA signatures Pathogens similar to nonpathogens complicating test development quotDirty world thousands of organism exist Microbial Forensic Technologies Technologies that identify the microorganism or toxins produced by microorganism Microbiological growth Microscopy ELISA Mass Spectrometry RealTime PCR assays Technologies that characterize the microorganism DNA Sequencing SNPs VNTRs Technologies that characterize how the microorganism was grown on what else might be present Microscopy Mass Spec Microbiology Future Challenges to Microbial Forensics Development and validation of new methods for forensic microbiology DGeneral acceptance of methods via peer review publications Integration of both the IC and Law enforcement forensic communities DEducation of the quotvoicesquot and decision makers of both communities Lawyers judges analysts policy makers Developmentrecruitment of the next generation National Policy Concerns amp Definitions USG Policy for Attribution of Weapons of Mass Destruction Attacks Annex V to NSPD17HSPC4 Purpose The policy clarifies roles responsibilities and authorities of USG departments and agencies regarding the attribution of a WMD attack or attempted attack on the homeland US citizens or US interests abroad this policy also outlines the USG process for providing appropriate officials with timely authoritative assessments of the sources and perpetrators of a suspected or actual WMD attack WMD is quotany weapon or device that is intended or has the capability to cause death or serious bodily injury to a significant number of people through the release dissemination or impact of toxic or poisonous chemicals or their precursors a disease organism or radiation or radioactivity WMD Attribution is quotthe capability and process to identify the nature source perpetrator and pathway of an attempted or actual WMD attack This includes rapid and comprehensive coordination of intelligence reporting law enforcement information technical forensics information and other relevant data streams to evaluate adversaries capabilities resources supporters and modi operandi in the context of a recent completed or attempted WMD attacks Forensics is quotthe scientific analysis and characterization of nuclearradiological chemical or biological materials samples devices output signals andor debris resulting from the use or attempted use of such material People responsible 0 Director of National Intelligence Attorney General Secretary of Homeland Security Secretary of State Secretary of Defense Secretary of Health and Human Services The Nuclear Regulatory Commission Environmental Protection Agency Brief History of Bio warfare 6th century BC Assyrians in Mesopotamia poisoned wells of enemies with rye ergot fungus Claivceps purpurea mycotoxins D1346 Tartars catapulted dead bodies infected with plague Yersinia pestis into the Black Sea port city of Kaffa Survivors may have been responsible for the second wave of Black Death D 1754 British General Jeffrey Amherst during the French amp Indian War reported to have distributed smallpox quotinfectedquot blankets to the Indians 1915 Germans during WWI infected the Russian livestock with Burkholdaria mallei a highly infectious bacteria that causes Glanders disease in horses and occasionally in man D1937 Japanese during WWII had Unit 731 facility in Manchuria Tested various organisms including anthrax on prisoners and the Chinese Japanese dropped bombs with Y pestis infected fleas on Chinese cities 1942 British test aerosolized anthrax on Gruinard Island near Scotland Island was decontaminated in 1986 1942 US begins offensive BW program at Camp Detrick MD D1969 President Nixon stops offensive BW program D1972 United States and several other countries sign the Biological Weapons Convention and destroy stockpile D1979 Sverdlovsk Russia has an accidental release of aerosolized anthrax from research facility D1980 s Russians attempted to use B mallei in Afghanastan D1991 Iraq s BW program discovered following Desert Storm US BW program 1942 Pres Roosevelt initiated a US offensive BW program with creation of War Reserve Service George W Merck in charge 1943 Camp Detrick becomes operational as the parent research and pilot plant center with about 4000 personnel 1960 s simulant testing conducted in New York San Francisco Washington DC to determine vulnerability of U S cities to covert BW attack 1969 President Nixon visited Ft Detrick on 25 November 1969 and announced a new national policy on BW quotThe US shall renounce the use of lethal biological agents and weapons and all other methods of biological research to defensive measures such as immunization and safety measures US Army Medical Research Institute of Infectious Diseases USAMRIID continued its mission to quotperform studies on the pathogenesis diagnosis prophylaxis treatment and epidemiology of naturally occurring infectious diseases of military importance with emphasis on problems associated with the medical defense against biological agents and on those microorganisms which require special containment facilities quot BW Agents of the US program Bacterial agents Bacillus anthracis Anthrax Franciscella tularensis Tularemia Brucella suis Brucellosis Coxiella burnetti Q fever Viral agents Venezuelan equine encephalitis virus VEE Yellow fever Toxins Botulinum Toxin Staphlococcal Enterotoxin Antiplant agents Pyricularia oryzae rice blast Puccinia graminis tritici wheat rust Simulants Bacillus globigii Bacillus subtilis Serratia marcescens Bioterrorism quotintentional or threatened use of bacteria fungi or toxins from living organisms to produce death or disease in humans animals and plants and involves quotintimidation of nations or people to accomplish political or social ends Bioterrorism Federal Laws Use of Weapons of Mass Destruction Statutes Title 18 Congress has enacted a number of statutes that provide criminal jurisdiction over the use of biological 175 chemical 2332c nuclear 831 and other weapons of mass destruction 2332a Biological Weapons Anti Terrorism Statute of 1989 BWAT Title 18 USC 175 178 DAmended 102401 to include Possession Without Legitimate Purpose as a Violation Public Health Security and Bioterrorism Preparedness and Response Act of 2002 Bioterrorism Act Weapons of mass destruction Any explosive or incendiary device as defined in Title 18 USC Section 921 bomb grenade rocket missile mine or otherdevicewitha chargeofmorethanfourounces DAny weapon that is designed or intended to cause death or serious bodily injury through the release dissemination or impact of toxic or poisonous chemicals or their precursors DAny weapon involving a disease organism DAny weapon that is designed to release radiation or adioactivity at a level dangerous to human life D vvru Diurugiucu Agent 0L lets ANY biological material capable of causing death disease or other biological malfunction in a human an animal a plant or another living organism deterioration of food water equipment supplies or material of any kind or harmful alteration of the environment Acts Knowingly develops produces stockpiles transfers acquires retains or possesses a biological agent toxin or delivery system for use as a weapon Or knowingly possesses any biological agent toxin or delivery system not reasonably justified by a prophylactic protective bonafide research or other peaceful purpose Agroterrorism The deliberate introduction use or threatened use of a chemical biological toxic radiological nuclear or explosive agent against one or more components of the food or agriculture sectors with the goal of causing mortality and morbidity generating fear precipitating economic loss or undermining sector stability and confidence in government Terrorist Interest in Agroterrorism A member of the Movement for Islamic Reform in posted an internet article in December 2003 stating that mad cow disease cost the United States 27 billion and this disease or other viruses could easily be transported to the United States He suggested that al Qa ida s goal of weakening the US economy could be achieved through several biological attacks including an agricultural disease a cattle disease a poultry disease and an epidemic disease ruuu 39clllu agriculture targets Farm to Fork cycle Livestock Crops Farm Workers Water supplies Livestock producers Food and agriculture transportation system Food processors and handler Processing facilities Restaurant grocery stores markets Agro Vulnerabilities Mother Nature provides many plant and animal diseases D Many foreign animal diseases Higth contagious Higth infectious Incubation period Helps spread outbreak before detection FMD in the UK spread for 3 weeks prior to detection DTypical livestock transportation can exacerbate spread DA very open system wide expanses of crops monocropping Potential amp Economic Impacts Economic effects oExport restrictions lost markets Direct costs treatment prophylaxis decontamination depopulation Lost revenues productivity wages Processing transportation distribution marketing tourism etc US Mad Cow Disease 23 December 2003 First US case only one cow Meats enttofoodandcommerciaprocessorsbeforediseasewasdetected Export bans South Korea resumed importation of US beef on 23 April 2007 Exports to 4 countries over 90 of US exported beef dropped from 3 billion 2003 to 467 million United Kingdom FMD Outbreak 2001 Cost estimated at 15biion Significant effects across multiple industries and economic sectors effecting the GDP for several years DTaiwan FMD Outbreak 1997 Caused country s share of worldwide pork exports to drop from 15 to none conomic Impacts E coli contaminated Spinach Sept 2006 DTargeting food supply results in Agro Vulnerabilities Many foreign animal diseases are endemic to regions where terrorists are known to reside Low cost little technical expertise required to acquireproduce Pathogens are easy to disseminate no need to weaponize safe to handle Recent events may inspire attacks DFood and agriculture are soft targets DLess morally objectionable than attacking people Select Agent Program The CDC HHS select agent program regulates the possession use and transfer of biological agents and toxins that could pose a severe threat to public health and safety 1996 Antiterrorism and effective Death Penalty Act Title V directs the Secretary of HHS to promulgate regulations identifying biological agents that pose a potential threat to public health and safety and governing their intentional or inadvertent transfer 2001 USA PATRIOT Act and the Bioterrorism Act expanded Select Agent Regulations List of select agents increased Lab personnel have to undergo a Security Risk Assessment SAR for access to Select Agents SAR s conducted by FBI Select Agent List Criteria 1 the effect on human health of exposure to the agent or toxin 2 degree of contagiousness of the agent or toxin and the methods by which the agent or toxin is transferred to humans 3 the availability and effectiveness of pharmacotherapies and immunizations to treat an prevent any illness resulting from infection by the agent or toxin and 4 any other criteria including the needs of children and other vulnerable population that the Secretary considers appropriate gDC list of Bioterrorism Pathogens LlSt includes organisms that pose a risk to national security because they D can be easily disseminated or transmitted from person to person result in high mortality rates and have the potential for major public health impact might cause public panic and social disruption and require special action for public health preparedness D Anthrax Bacillus anthracis D Botulism Clostridium botulinum toxin D Plague Yersinia pestis D Smallpox variola major D Tularemia Francisella tularensis D Viral hemorrhagic fevers filoviruses eg Ebola Marburg and arenaviruses eg Lassa Machupo C LlSt includes emerging pathogens that could be engineered for mass dissemination in the future because of availability ease of production and dissemination and potential for high morbidity and mortality rates and major health impact l l In n A A n A n n I IAL an I39uIln B LlSt includes organisms that are moderately easy to disseminate D result in moderate morbidity rates and low mortality rates D require specific enhancements of CDC39s diagnostic capacity and enhanced disease surveillance Brucellosis BruceIa species B Epsilon toxin of Clostridium perfringens Food safety threats eg Salmonella species Escherichia coli 01572H7 Shigela D Glanders Burkholderia mallei D Melioidosis Burkholderia pseudomalei D Psittacosis Chlamydia psittaci D Q fever Coxiella burnetii Ricin toxin from Ricinus communis castor beans U Staphylococcal enterotoxin B U Typhus virus Rickettsia prowazekii U Viral encephalitis alphaviruses eg Venezuelan equine encephalitis eastern equine encephalitis western equine encephalitis n Ala l39or ca Fo hI hroac In a il rin rhnlorno Biological Terrorism Rajneesh Foundation 1984Biological Attack in the Dalles Oregon Salmonella Contamination of 8 Restaurant Salad Bars 751 Citizens Affected Attempt to Manipulate Local Election Aum Shinrikyo 1984 Shoko Asahara started a yogameditation group 1989 designated quotreligion of the elite DResponsible for murdered anticult attorney Sakamoto in 1989 D1995 Sarin attacks in Tokyo subway Killed 12 injured 50 affected 980 DAsara was arrested convicted of murder amp sentenced to death DSearches of compound revealed laboratories and equipment interest in anthrax C botulinum sarin VX 1993 Aum Shinrikyo Anthrax June July 1993 Tokyo D Reports of quotFoul Odors to local Environmental Health Officials D quotGelatin like oily gray black fluid from the mist from the bld cooling towers DNo entry granted external site visit samples taken D 1999 liquid samples tested positive for B anthracis Stern 34F2 strainoften used commercially in Japan to vaccinate animals against anthrax Non Pathogenic Strain Amerithrax Investigation Involves the distribution of anthrax via the US Postal Service 2 sets of letters were mailed on different days D22 human cases of anthrax Inhalation and cutaneous D5 deaths Dramatic changes to USG policies programs institutions and laws Seven year investigation DDevelopment of microbial forensic techniques AmerithraX Investigation First Strike Robert Stevens First victim and first indication of the attack Lived in Boca RatonFL Editor at tabloid quotThe Sun Became ill Sept30th died Oct5 of inhalation anthrax Anthrax detected at AMI office building Coworkers also exposed to anthrax Source believed to be a letter mailed to the office This letter was never recovered Similarities to the New York Post and NBC letters DReference to 911 B Death to America Death to Israel Allah is great D Differences Second Strike Daschle letter October 9 2001 Letter mailed to Senator Daschle Opened by a staff member in the Hart Building on Oct 15 40 staffers were present 28 were exposed Hart Building was closed 4 Washington DC postal workers were exposed at the Brentwood mail handling facility Two later died of inhalation anthrax on Oct 22 Anthrax contamination found at buildings processing mail for CIA White House and Supreme Court Letter mailed to Senator Leahy was not delivered Mail from Brentwood was sequestered and searched Leahy letter found Nov 11th D Postmarked from 4th grade Greendale Elementary School Franklin Park NJ D Contained the warning quotyou cannot stop us We have this anthrax You die now Leads in Anthrax evidence M39Cmb39al Forens39cs Nonmotile Grampositive endospore forming bacterium Identified by Robert Koch in 1876 first organism to be linked Relatively young field of forensics disease Koch39s postmates D Rely on traditional microbiology Causative agent of anthrax D Utilize existing genetic typing Vegetative bacmus rapid growth 1520 min doubling time D D Develop new approaches noninfectious techniques D Lucky that it was anthrax because research had been done on this mm Metabolically dormant D Stable for decades D Resistant to UV heat desiccation chemical organism D infectious particle AnthraX known population genetics Closely related to Bacillus cereus Some strains of Bacillus anthracis has been sequenced entirely Dr Paul Keim Northern Arizona UniV had done extensive work DNA typing of different strains of anthrax VNTR and SNP typing techniques Determined it was Ames strain anthraX AnthraX Ames Strain Amerithrax Microbiology Ames straIn of anthrax used In all letters Samples from the letters were cultured at USAIVIRIID Ames first isolated quotdiscoveredquot in 1981 in D h ff r 39 39 39 e e mInor quotvariantquot type of colonies Chief Use Laboratory Strain Challenge strain in anthrax vaccine research and development Ames was never used in the US or Soviet offensive biological weapons programs Whole genome sequence available Amerithrax DNA Sequencing Sequencing done by Dr Jacque Ravel and Dr Claire Frasier TIGR Maryland Inst Genomic Research DSequences were compared D from the letters U The original ancestral Ames still located at USAMRIID U Wildtype WTand variants No sequence difference between the WT anthrax from the letters and the ancestral Ames strain D Sequence differences of the variants from the WT DThese contaminants could help sort out the various quotbatchesquot of Ames used by laboratories around the world AmerithraX Microbial Forensics Spores from the letter contained quotva riants B DNA Sequencing of variants found differences D Assays were developed and four different labs conducted the subsequent analyses D All Ames isolates were obtained and screened 1070 samples from different laboratories around the world D Eight isolates had the four variants D RMR 1029 from USAMRID Amerithrax RMR 1029 8 samples contained all 4 genetic mutants found in the anthrax letters D All 8 samples were historically linked to one flask of purified spores D RMR 1029 spores were produced and used for vaccine testing at USAMRIID D The 1 liter spore flask called RMR 1029 contained concentrated spores from 34 production lots gt160 liters of Ba12 fermentor and 22 multiple flask cultures Amerithrax Perpetrator Dr Bruce Ivins PhD Microbiologist who at USAMRIID for 36 years D Interest in anthrax and development of vaccine following Sverdlovsk anthrax release Coinventor for two anthrax vaccine patents DResponsible for RMR1029 as he used it in vaccine challenge experiments Worked on Amerithrax evidentiary samples from letters Died on July 29 apparent suicide from overdose of Tylenol National RampD Strategy Purpose to guide and focus the research efforts of the US Government to advance the discipline of microbial forensics and provide the nation with the most scientifically sound and statistically defensible capability to provide scientific data to support attribution investigations of a potential or actual biological attack Strategy was developed by a joint committee with representatives from the DNI DHS DOJ DOD and HHS Issued Dec 2009 by the Executive Office of the President via the National Science and Technology Council
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