BiologicalEvidence Screening&Serology

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BiologicalEvidenceDetection,CollectionandSerology

Thankyouforyourparticipationinthistrainingmodule.Themoduleisavailableonlineandisavailable24hoursa dayand7daysaweekfor30daysafterenrollment.Thecourseisselfpaced,whichallowsyoutocontrolthe progressionofthetraining.Youmaypauseandresumeasoftenasneeded.Simplycloseyourbrowsertoquitand loginathttp://www.forensictrainingnetwork.com/_pages/stsignin.htmltoresume.

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Instructions
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CourseContentandQualifications

ThiscoursewasdevelopedbyexperiencedforensicDNAscientistsJaimeHandelsmanandCatherineCaballero. Theirbiographiesareprovidedbelow. JaimeHandelsman,B.H.S.

JaimeHandelsmaniscurrentlycofounderandownerofForensicTrainingNetworkandisresponsiblefor developinganddeliveringonlineandonsiteforensicDNAtrainingaswellaspartneringwithexpertstooffer onlinetrainingforvariousotherforensicdisciplines.Jaimehasbeendevelopinganddeliveringtrainingcourses, lecturesandworkshopsonthelatestforensicDNAtechnologiessince2001.Shehastrainedhundredsofforensic analystsfromcrimelaboratoriesacrosstheUnitedStatesandhasgivennumeroustechnicalpresentationsat industrymeetingsandconferences.Shealsoparticipatedinthedevelopmentandsupportofvariouscommercial forensicDNAtestingproductsprovidedbyAppliedBiosystemswheresheworkedaspartoftheirHuman Identification(HID)teamfrom2001until2007.

BeforeAppliedBiosystems,Ms.HandelsmanwasemployedbyTheBodeTechnologyGroupinVirginiatodevelop andimplementaqualitycontrolprogramandmanagelaboratorytechnicians.Jaimebeganhercareerinforensic scienceattheArmedForcesDNAIdentificationLaboratory(AFDIL)inRockville,Marylandwhereshereceived traininginmtDNAandSTRanalysis,playedasignificantroleinthelaboratorysqualitycontrolprogramand participatedinotherforensicresearchprojects,suchasautomation.Jaimesuccessfullycompletedadvanced geneticandforensicbiologycoursesaspartoftheForensicMolecularBiologygraduateprogramattheGeorge WashingtonUniversityinWashington,D.C.inadditiontoearningaBachelorsofHealthSciencedegreefromthe UniversityofMiamiinCoralGables,Florida. jaime@forensictrainingnetwork.com|18669636835x201 CatherineCaballero,B.S.

CatherineCaballerohasbeendedicatedtothefieldofforensicsciencesince1994.Herpresenceinthecommunity isstrongasaresultofherextensiveexperienceworkingwithforensicscientistsaroundtheworld.Asacofounder andownerofForensicTrainingNetwork,sheprovidestrainingtotheforensiccommunitybydevelopingonline trainingmodulesanddeliveringonsitetrainings.

Previously,sheworkedforAppliedBiosystemswhereherprimaryresponsibilitywastoprovidetrainingand troubleshootingsupporttoforensicDNAscientists.Shealsogainedextensiveexperiencedevelopinganddelivering handsontrainingprograms,conductingtechnicalworkshopsandpresentingatseminarsandtradeshows. Catherineisconsideredtobealeaderinthisarenaandisamentortomanyofhercolleagues.

CatherineworkedfortheTexasDepartmentofPublicSafetyCrimeLabduringtheyearsinwhichDNAtechnology wasfirstadoptedbythestateofTexas.ShewascreditedwithestablishingtheDNAlabinCorpusChristi,validating andimplementingSTRtechnologyinHouston,andprovidinghandsontrainingforhercolleaguesthroughoutthe state.Inaddition,CatherineconductedcrimesceneinvestigationswiththeTexasRangersandperformed caseworkanalysisincludingserology,DNAanalysis,andcourttestimony.Duringhercollegeyears,Catherine internedattheCaliforniaCriminalisticsInstituteandworkedfortwoprivatelabs.ShereceivedaBachelorsof ScienceinForensicSciencewithaminorinChemistryfromtheCaliforniaStateUniversity,Sacramento. catherine@forensictrainingnetwork.com|18669636835x202

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BiologicalEvidenceDetection,CollectionandSerology

ThistrainingmoduleincludestechnicalinputandreviewbyforensicexpertPatriciaGraham.Patriciahas spentthelast30yearsinthefieldofforensicDNAandserologyandhasrecentlyretiredfromtheTexas Dept.ofPublicSafety.ShewasthemanagerandDNAtechnicalleaderforthecrimelabinCorpus Christi,TX.DuringheryearswiththestateofTexas,Patwasresponsibleforproceduremanuals, training,DNAcasework,crimescenes,andkeepingupwithaccreditationstandards.

ModuleDuration

Thismoduleisdesignedtobecompletedfromstarttofinishin1.5hours.However,timesmayvary sincethetrainingisselfpaced.

Objectives

TheultimategoalofanalyzingbiologicalevidenceistodeterminethecontributorbasedonDNAtesting, butwemustfirstcollect,detectandidentifythebiologicalevidence.Thegoalofthismoduleisto provideathoroughunderstandingofthetypesofbiologicalevidenceencounteredinforensic investigations,themethodsusedtodetectbiologicalevidence,thestepstakeninthelaboratoryto processbiologicalevidenceandthevarioustypesofserologytestsusedtoidentifyblood,semenand saliva.

Agenda

TypesofBiologicalEvidence BiologicalEvidenceConsiderations CollectingBiologicalEvidence ChainofCustody BiologicalEvidenceProcessing&Considerations EvidenceScreening EvolutionofSerologyTesting SerologyTesting Blood Semen Saliva

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Section1:TypesofBiologicalEvidence
LocardsExchangePrinciple
Whereverhesteps,whateverhetouches,whateverheleaves,evenunconsciously,willserveas asilentwitnessagainsthim.Notonlyhisfingerprintsorhisfootprints,buthishair,thefibers fromhisclothes,theglasshebreaks,thetoolmarkheleaves,thepainthescratches,theblood orsemenhedepositsorcollects.Alloftheseandmore,bearmutewitnessagainsthim.Thisis evidencethatdoesnotforget.Itisnotconfusedbytheexcitementofthemoment.Itisnot absentbecausehumanwitnessesare.Itisfactualevidence.Physicalevidencecannotbewrong, itcannotperjureitself,itcannotbewhollyabsent.Onlyhumanfailuretofindit,studyand understandit,candiminishitsvalue. EdmondLocard,1910

ExchangeTheory
Whentwosurfacescomeincontactwitheachother,partial exchangeortransferofonematerialtotheotheroccurs Basisforcrimesceneinvestigation Evidence(materialtransfer)overlookedandnotcollectedduring theinitialinvestigationmightnotberetrievableand/oradmissible incourt

TypesofTransfer
PrimaryTransferthedirecttransferofDNAfromonepersontoanobjectoranother person SecondaryTransferDNAdepositedononeitemorpersonistransferredtoasecond objectorperson TertiaryTransferDNAontheseconditemorpersonistransferredtoathirdobjector person

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TypesofBiologicalEvidence

Evidenceisusedincriminalinvestigationstohelpproveordisproveanindividual'sinvolvementina crime.Evidencemaybe______________________or__________________________.Biological evidenceisdefinedas_____________________evidenceoriginatingfroma _______________________________,suchasa______________.Therearevarioustypesofbiological evidenceandwithineachtypetheremaybemorethanoneform.Thetype,form,quantityandquality ofthebiologicalevidencewilldeterminethe_________________________and _________________________usedtocollectandprocessit.

(Enterabriefdescriptionforeachtypeofbiologicalevidencepresented)

Type
Blood

Description

Semen

Saliva

Bones and Teeth

Hair

Epithelial Cells

Tissue

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Section2:BiologicalEvidenceConsiderations
ChallengeswithBiologicalEvidence
MixedwithmoleculesthatpreventafullDNAprofilefrombeingobtained Biologicalmaterialcollectedmaybeamixturefromtwoormorehumansources Limitedsamplequantity ExposedtoharshenvironmentalconditionswhichcausestheDNAtodegrade

TTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTAC

TTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTACTTAC

Contamination

CausesofDegradation
Samplesmaybeexposedpriortooraftercollectiontothefollowing: - ExcessiveHeat - Environmentalconditions Sunlight Rain Soils,bacteriaandfungi - Chemicalexposure Bleach,detergents - UVlight Fromsunlight SomelaboratorieswilltreatworkspacewithUVlightaspartoftheir cleaningandsterilizationprotocols Propercollectionandstoragewillhelppreventdegradation

Contamination
AdventitioustransferofDNA Transferfrom - Evidencetoevidence - Persontoevidence Canoccuratcrimesceneorinlaboratory Laboratoriesmustrecordoccurrencesandcorrectiveactiontaken Questionsintegrityofevidence

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TypesofContamination
_______________________ - Oftenduetopoorhandlingproceduresbyconsumablesuppliers _______________________ - Transferofevidencetoevidence - OccursduringinitialevidenceexaminationortheDNAextraction _______________________ - Transferfromselftoevidence - OccursduringtheinitialevidenceexaminationorDNAextraction

ContaminationPrecautions
Gloves Footsiesorshoeprotection Hairtiedbackorhat/capworn DoNOTtouchfaceormouth DoNOTcough,sneezeortalkoverevidence Decontaminateequipment,toolsandworkspace Handleeachevidenceitemseparately

LaboratoryContaminationPrecautions
Separatelabspace/roomfordifferentanalyticalsteps - PrePCRlab EvidenceProcessingandScreening Extraction Quantification STRAmplificationSetUp - PostPCRlab PCRInstrumentOperation(STRamplification) CapillaryElectrophoresis Performmostsensitiveanalyticalproceduresinlaminarflowhoods Disposableplastics

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Section3:CollectingBiologicalEvidence
CollectingBiologicalEvidence
Transportableobjectsshouldbecollectedintheirentirety - Fewermanipulations - Lesschancefor___________________________ - Unstainedportionofobjectoftentested(substratecontrol) Ifnotpossible,thebiologicalmaterialshouldbe - cut - Scraped - tapelifted - transferredusingmoiststerileswab

BiologicalEvidencePackaging
Bloodandbodyfluidsshouldneverbepackaged_________ - Avoidscontamination - Bacteriagrowthcausesdegradation - Airdryatroomtemperatureinasterileandsecurearea - Usingexcessiveheat(suchashairdryers)andUVexposure fromsunlightcancauseDNAtodegrade Driedstainsandevidenceitemsshouldbecollectedinto________ bindlesorenvelopesandputinto__________evidencebags - Neveruseplastic - Exposuretoair(breathing)iscriticaltopreventbacterial/fungalgrowth Eachstain/itemshouldbecollectedandpackagedseparately

AdditionalSampleCollections
____________________Sample - Collectedfromtheindividualsreportedlynotinvolvedincrime - PresenceofDNAmightbedetectedandisexpected - Collectedfromindividualswithaccesstothecrimescene - Mayhelpdeterminesourceofcontamination - Preventsevaluatingunknownprofiles ____________________Sample - Collectedfromanyoneinvolveddirectlyinthecrime Samplescollectedbyswabbinginsideofcheek(buccalcells)
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CollectingDNAfromSuspects
UnconstitutionaltoobtainDNAsamplewithoutprobablecauseorconsent LegalcollectionofDNAsample - Searchwarrant - Courtorder - Legislativemandate - Voluntaryconsent ____________________sample - Leftbysuspectsinpublicplaces i.e.,cigarettebutt,drinkingcuporchewinggum - Noinvasionofprivacyorviolationofconstitutionalrights - Chainofcustodyandsampleauthenticationarecritical

Section4:ChainofCustody

WhatisChainof Chain of Custody (COC) is a written or an electronic record documenting the Custody(COC)? following as it relates to physical evidence:

the ____________________________ the __________________________ and ___________________________ the ____________________________ the ____________________________ the ____________________________ the ____________________________

Purposeof Chainof Custody

Evidence must be preserved and protected to maintain its ___________________________________ _______________________________. Chain of Custody is required to authenticate evidence in a court of law. It provides proof that the evidence item(s) has not been altered, substituted, or contaminated. Documenting the location, possession and handling of evidence at all times provides assurance that the item collected at the crime scene, tested in the laboratory and presented in court are one in the same. If the Chain of Custody is broken, the integrity of the evidence comes into question.

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Establishing Chainof Custody

All physical evidence items must be collected and packaged appropriately. The packaged evidence is identified with the case number, item number, initials of the individual collecting the item and the date/time of collection. In addition, all packaged items are sealed, preferably with crime scene evidence tape. Crime scene evidence tape is specifically designed to make obvious any attempts at tape removal or package entry. A proper evidence seal includes the initials of the individual placing the seal on the package. Initials are clearly written across the tape in permanent ink. Some agencies will also note the date of collection on this evidence seal. When evidence is collected during a crime scene investigation, a Crime Scene Evidence Collection Log is created to document the retrieval of each item. This log includes the following information:

____________________________ ____________________________ ____________________________

____________________________ ____________________________

LabChainof CustodyForm General Information

Upon submission of physical evidence to the crime laboratory, an accompanying Chain of Custody form is required. Here is an example of the general case information to include: ____________________________ ____________________________ ____________________________ ____________________________ ____________________________ ____________________________

LabChainof CustodyForm Item Submission

Upon submission of physical evidence to the crime laboratory, an accompanying Evidence Submission form is required. The following information is typically included on an evidence submission form:

____________________________ ____________________________ ____________________________

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LabChainof CustodyForm Transferof Evidence

Any time a physical evidence item is transferred from one agent or agency to another, it must be documented in order maintain its legal integrity. The information required includes: ____________________________ ____________________________ ____________________________ ____________________of individual ____________ possession of the item ____________________of individual ____________ possession of the item

Maintaining Chainof Custodyinthe Lab

Once physical evidence is received by a crime laboratory, it is stored in a _______________________ with ________________________ such as an evidence storage room or locker. For any laboratory analysis to be performed, the scientist must take possession of the evidence and become responsible for its custody and care. All evidence examinations and laboratory testing is documented in detail. The scientist in possession of the evidence will either maintain custody in a secure working area or in a short term storage area or locker when not performing direct examinations. At times, evidence requires the analyses of multiple laboratory sections such as DNA and fingerprints. In this event, the subsequent analyst will retrieve the evidence from the evidence storage room after it has been returned or evidence can be transferred directly from one analyst to another. Every instance of evidence transfer is documented to maintain the internal Chain of Custody.

Chainof Custody Considerations

There are a couple of additional factors to consider with respect to Chain of Custody for physical evidence. Limiting the number of individuals handling and possessing the physical evidence is preferred. The process of introducing the Chain of Custody into a court of law requires that every individual testify to their involvement with the evidence. In addition, minimizing the chances of evidence mis-handling and contamination is best accomplished by limiting the number of individuals in the Chain of Custody. Anytime a sealed package of evidence is opened, it is strongly recommended that the point of entry be a different and unique location. If at all possible, the previous seal should be kept intact. Once evidence examination is complete, the package point of entry needs to be properly sealed to include the initials of the handler written across the evidence tape in permanent ink. It is strongly recommended that the date also be documented across the evidence tape.

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Section5:EvidenceProcessing&Considerations
EvidenceAccessioning
Evidenceisloggedanddocumentedoncesubmittedtothelaboratory Eachevidenceitemisassignedanumber Oncesamplesaretakenfromapieceofevidence,samplenumbersareassigned Namingconventionandnumberingsystemsvaryamonglaboratories

GeneralAnalyticalWorkflow
Teststoindicate/identifytypeofbiologicalfluid andspecies Visualexamination

IndividualIdentification

AnalyticalTests
______________________Tests - Positiveresultsindicatethatthesampleisprobablythesubstancebeingtested for - Lacksspecificitysinceothersubstancescangiveapositiveresult ______________________Tests - Positiveresultsprovidesconclusiveevidencethatthesampleisthesubstance beingtestedfor - Offersspecificitysincenoothersubstancecangiveapositiveresult

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EvidenceStainSampling
Stainsshouldneverbetesteddirectly Samplingshouldbetaken - Prevents__________________________ - Protectsremainingstainforfurthertesting Smallcutting,dryswabbing,rubbingorscrapingcanberemovedfromstainfortesting Transfermethodsusingsterilewaterinclude - ___________________ofsample - Overlaywithfilterpaper

ConsumptionofEvidence
Untestedportionoftheevidencestainorreferencesampleshouldbepreservedfor retestingwhen: - theinitialtestresultsareinconclusive - theresultsindicatecontamination - thedefendantexercisesrighttodoso - thecourtrequestsasecondopinion - newtechnologyemerges Ifentiresampleneedstobeconsumedforsufficientresults,laboratoryprotocolsmust bereferenced - Mightrequireacourtordertoproceedwithtesting - Awitnessmightberequestedtoshadowthetestingprocess

StorageofEvidence
Shorttermstorage - _____________________________ Stainsonsmoothandnonporoussurfaces Stainsrequiringfingerprintanalysis Minimizesissueswith___________________________ Longtermstorage - Freezer Optimalstorageconditionsforbiologicalmaterial - Refrigerator Liquidbloodstoredinvialsthatmaybreakiffrozen Wetevidencestainsmustbe___________beforestoring

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Section6:EvidenceScreening
Screening
Carefullyexamineevidenceforbiologicalstains - Strongwhitelight - UVlight - ALS - Microscopes - Magnifyingglasses Carefulnotesandphotographsaretakenduringtheprocess

DetectingBloodstains
Strongwhitelight(LEDlightpreferred)forbloodstain(s) visibletothenakedeye - Freshbloodwillappeardarkred - Colorofblooddarkensasitdries;oldstainscan appeardarkbrownorblack Bloodstainsnotvisibletothenakedeye - UVlight - Chemiluminescentorfluorescentchemicalreactions

Chemiluminescent&FluorescentDetection
Detect_______________amountsofbloodandcleanedbloodstains Luminol,fluorescein,andBlueStar Sprayedoversuspectedarea Emitsa____________________to__________________glow Consideredapresumptivetestduetopositiveresultswithmetals,bleach,treated lumberandanimalfat Protectionshouldbeworn - maycauseeyeandskinirritation - maycauserespiratoryanddigestivetractirritation Catalytictestsbasedon_________________reaction

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CatalyticReactions
_________________=achemicalthatwillinitiateoraccelerateareaction - Hemoglobininbloodcontains____________ - Ironactsasacatalystforoxidationreactions Oxidationreaction:reactioninvolvingthelossof________________byamolecule - _____________________________typicallyusedastheoxidant - Newoxidizedchemicalisproducedresultingincolorchangeorreleaseoflight

Luminol
Visualizedinavery___________environment ChemiluminescentReaction - Originalmoleculesinthereactionhavemoreenergythantheproductsofthe reaction - Extraenergycreatedisreleasedintheformofvisiblebluewhitelight ChemicalReaction - Luminolpowderismixedwithhydrogenperoxide - Ironinhemoglobinactsasacatalystfortheoxidationreaction - Luminollosesnitrogenandhydrogenatomsandgainsoxygen atomswhichresultsinelectronexcitation

FluoresceinandBlueStar
Fluorescein - Combinedwithoxidantandsprayedoverarea - Lightemittedasenergyislostinthereactionresultinginfluorescence - VisualizationwithUVlightinadarkenvironment - Effectiveon_________________surfacesduetopresenceofthickener BlueStarForensic - Luminescentreactiondoesnotrequiretotaldarknessforviewing - Fastdissolvingtablet

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DetectingOtherBiologicalEvidence
Visualexaminationwithwhitelight - Offwhitetoyellowishstainsmightbevisible - Slighttexturedifferencesduetohardenedstainsmightbevisible UVlight ALS(A__________L_________S__________)forbiologicalstainsthatarenotvisibleto thenakedeye

AlternateLightSource(ALS)
Usesdifferent________________and___________ toenhancestaindetection - ALShastheabilitytoadjusttheinput excitation wavelengthandtheoutputemission spectrumorrange - Differenttypesofbiologicalfluidswillhave optimalwavelengthrangesfordetection - Goalistoachievethegreatest_____________ betweenbiologicalfluidandsubstrate Barrier_______________ - Protectionfrompowerfullighttopreventanydamagetotheeye - Manufacturedwith_______(orange,yelloworred)toblockextraneousemitted wavelengthsandenableviewingoffluorescentemissionsbythehumaneye

Section7:EvolutionofSerology
(Enterthemajorevent/technologyintroducedduringeachyearandabriefdescription)

Introduction _______________ is the scientific study of blood and its properties. Current forensic

serology testing involves the use of chemicals to confirm a substance is biological in nature and/or determine the type of biological material present (i.e., blood, semen, saliva, etc.)

1839

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BiologicalEvidenceDetection,CollectionandSerology

1853

1863

1901

1925

1928

1931

1938

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BiologicalEvidenceDetection,CollectionandSerology

1945

1960s

1967

1986

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Section8:SerologyTestingBlood
BloodstainAnalysis GeneralWorkflow AlternateWorkflow1 AlternateWorkflow2

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PresumptiveBloodTesting

Introduction

While Luminol, florescein and BlueStar are used to detect blood stains not visible to the naked eye, they are also considered presumptive blood tests which produce a release of light from an oxidation reaction. Presented here are additional presumptive blood tests currently utilized in forensic science that produce a color change from an oxidation reaction. As previously mentioned, these tests are not specific for blood. Other catalysts and chemical oxidants might be present such as metals and bleach which could cause a false positive result for blood. Plant and vegetable peroxidases such as in horseradish, garlic, cabbage, and tomato may give false positive reactions for blood. In addition, other substances of animal origin could result in positive reactions. These catalytic tests are performed by mixing a specific reagent with an oxidant and exposing it to a sampling of the stain in question. Most catalytic test results are read immediately and are not considered valid after about a minute.

LCV&LMG

Leucocrystal Violet (LCV) does not have a widespread use in serology crime laboratories, but it has been adopted to aid in the detection of bloody fingerprints, footprints and shoe prints in the laboratory and at crime scenes. When LCV and hydrogen peroxide come into contact with the hemoglobin in blood, a catalytic reaction occurs and the solution turns a _______________________. Leucomalachite Green (LMG) reacts with hemoglobin in blood in a similar fashion to produce a __________________________.

Phenolphthalin Phenolphthalin is a commonly used forensic presumptive blood test and is also
known as the Kastle-Meyer test. A color change from clear to ________________ provides a positive result with moderately low sensitivity.

TMB

Tetramethylbenzidine (TMB) is another commonly used forensic presumptive blood test. A color change to ___________________________________ provides a positive result with moderate sensitivity. The ____________________________ product developed by Bayer consists of disposable test strips manufactured with TMB reagent affixed to test squares. Once a sample of the stain has been collected onto a swab or filter, the swab or filter is rubbed onto the test square. A color change is compared to a result chart provided with the product to determine a positive result. ________________________ manufactured by ODV Inc, also utilizes TMB. It is a selfcontained, disposable tube with a swab for stain collection and reagent ampoules. Using sterile water, the swab is used to sample the stain and is inserted into the tube. The two ampoules are broken to allow reagent contact with the stain on the swab. A blue/green color provides a presumptive positive result for blood.

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Immunology
____________________(Ag) - partsofforeignmatter(suchasharmfulbacteriaorviruses)inthebodythat triggersanimmuneresponse _____________________(Ab) - proteinmanufacturedtofightoffantigens - chemicallyrecognizedtoattachtoantigens,inactivatethemandberemoved fromthebody Whenantibodiesandantigensattach,anAgAblatticeforms(precipitinreaction)

TypesofAntibodies
Whatisamonoclonalantihumanhemoglobinantibody? - monoclonal singlespecificity - allbuiltalikeandmanufacturedbyasingleclone antihumanhemoglobin - specificityforhumanhemoglobin Polyclonalantibodiesareantibodiesthatarederivedfrommanydifferentcellsorcell linesandareamixtureofmanydifferentspecificities

AntigenAntibodyTesting
Sample(containingantigens)isaddedintoonewell Antibodiesaddedintosecondwell Outwarddiffusionofantigensandantibodies _____________________formswhereAgAb(antigenantibody)complexforms

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Biolog gicalEvidenceDe etection,Collect tionandSerolog gy

ABAcardHemaTr d raceTestSt trips

________ __________ ___________ _Test

Preloadedin ntothesamp plewellare:

1.Monoclo onalantihum manhemoglo obinantibod diesand 2.Dyetagg gedantibodiesasaninte ernalcontrol l

Whensamp plecontainin nghuman__ ___________ ________isaddedanAgAb complexisf formed.

Polyclonala antihumanhemoglobina antibodiesareattachedtothearea labeled___ _____.Ano othersetofa antibodiesfo ortheintern nalcontrol(IC)is attachedtothearealab beled_____ ____.

WhentheAgAbcomple W exandICmo oleculesmig grateintothe eimmobilize ed antibodie es,________ __coloredb bandswillap ppearinthei irrespective elocations. - TheTwilla appearduet toformation nofAgAbAg - TheCwilla appearduet todyeaccum mulation

raceTestR Results HemaTr

Hemoglobinc H concentratio onof0.05ug/mLisnecessarytoobtainaposit tiveresult Purposeofin nternalcontr rolistodemonstratetha atthetestisworkingpro operly T Tcanreact ttohuman,h higherprima ate,andferr retblood

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HexagonOBTITest
Humanhemoglobininthesampleformsacomplexwithmonoclonal antihumanhemoglobinantibodieslabeledwitha__________dye Thecomplexmigratestothetestzoneandiscapturedbyan immobilizedsecondantibody Apositiveresultoccurswithabluetestline Antimouseantibodiesareutilizedforthepositivecontrol Positivetestresultcanbeattributedtohuman,higherprimate, ferretandskunkblood

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BiologicalEvidenceDetection,CollectionandSerology

Section9:SerologyTestingSemen SemenStainAnalysis

GeneralWorkflow

AlternateWorkflow1

AlternateWorkflow2

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AcidPhosphatase(AP)Testing
Enzymesecretedbytheprostateglandintoseminalfluid SemencontainsAPinamountsupto400timesgreaterthanotherbodyfluids _____________________________forsemen - Strongpositivetestresultindicatesthepossibilityofthepresenceofsemen APTestChemicalReaction - AcidicsolutionofsodiumalphanapthylphosphateandFastBlueB(FBB)dyeis sprayedordroppedonswaborfilterpaper - InthepresenceofAP,acomponentofsodiumalphanapthylphosphateiscleaved andbindstoFBB - A_______________todark_____________colorindicatesapositiveresultfor thepresenceofAP

Spermatazoa
Microscopicidentificationofspermatazoaservesasa___________________testforthe presenceofsemen Spermatazoaarethegametecellsfoundinsemen Humanspermhasuniquephysicalmorphology

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MicroscopicSpermIdentification
Confirmationcanbeachievedwithatleastone - intactspermatozoanwithtail - spermatozoanhead Microscopicslidepreparation - Duringsexualassaultexaminationofvictim Evidenceswabs(vaginal,rectal,oral,etc)fromthevictimaresmeared ontoamicroscopicglassslide - Slidesmaybecreatedfromanyadditionalevidenceswabsandstainslater Stainisdissolvedinsterilewater Portionofthepelletisplacedonamicroscopeslide

VisualEnhancementofSperm
_______________________Stain(KernechtrotPicroindigocarmine) - Kernechtrot(NuclearFastRed)solutionstainsthe spermheadareddishhue - Picroindigocarminesolutionstainsthespermtailsa greenishhue SpermHYLITERKitfromIndependentForensics - 4',6diamidino2phenylindole(DAPI)usedto fluorescentlylabelDNA - Fluorescentlylabeledantibodiesthatbindtohuman spermheadsusingFluoresceinisothiocyantate(FITC) - VisualizationrequirestheuseofamicroscopewithDAPIandFITCfilters

SpermDetectionTimes

Saferstein,R.(1988)ForensicScienceHandbook,VolumeII.EnglewoodCliffs,NewJersey:PrenticeHall,Inc.

Note!Timesmaybehighlyvariable

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SpermCountClassification
Approximatenumberofaveragespermdetected permicroscopefieldofview Providesgeneralassessmentof______________ forDNAanalysis Lackofspermdoesnotindicatelackofsemen

ProstateSpecificAntigen(PSAorp30)Testing
ProstateSpecificAntigen(PSAorp30)isahumanproteinproducedinlargequantities bytheprostategland Testreliesonantigen(PSA)andantibodyinteraction Valuableforidentifyingsemencontributedby_____________or______________ individuals Onceconsideredonlyapresumptivetestduetoobservedcrossreactivitywithother bodyfluidsusingolderdetectionmethods - Ouchterlony - Crossoverelectrophoresis - Rocketimmunoelectrophoresis - Radialimmunodiffusion - ELISA(enzymelinkedimmunosorbentassay) ABAcardp30TestStripsconsidered___________________forthepresenceofsemen Positiveresultnotobtainedfromtraceamountsthatmightbefoundinotherbodyfluids Nocrossreactivityobserved

ABAcardp30TestStrips
ReactionpremiseisthesameasHemaTraceTestStrips UtilizesmonoclonalantihumanPSAantibodiesinsamplewell PolyclonalantihumanPSAantibodiesareattachedtoastripofthe membraneinthetestresultarealabeledT Internalpositivecontrolpresent APSAconcentrationof4ng/mLorgreaterisnecessarytoobtaina positiveresult

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SemenogelinDetection
Semenogelinisaproteinfoundinhumansemenandcanbetestedto_______________ thepresenceofsemen RSIDSemenfromIndependentForensics - Utilizestwomonoclonalantibodiesspecificforsemenogelininsamplewell - Antisemenogelinantibodiesareattachedtoastripofthemembraneinthetest resultarealabeledT - Internalpositivecontrolpresent

Section10:SerologyTestingSaliva

Amylase
Presenceistestedtodetectsaliva Enzymethatbreaksdownstarch Pancreaticamylase - Semen - Feces - Vaginalsecretions Salivaryamylase - Saliva - Perspiration - Breastmilk

AmylasePresumptiveTests
StarchIodineTest - Iodineandstarchreagentis______________ - Amylasewillbreakdownthestarchandthebluecolorwillfade PhadebasReagent - Starchlinkedtoacoloreddye - Amylasewillbreakdownthestarchandreleasethedyeresultingina________ color SALIgAESalivaTest - Atubecontainingclearsolutionturns____________inthepresenceofsalivary amylase - Specificforsalivaryamylase

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AmylaseConfirmatoryTest
RSIDSaliva Utilizestwoantisalivaryamylasemonoclonalantibodiesinteststripformat Internalpositivecontrolpresent DetectionlimitensuressufficientamountsforfurtherDNAtesting Manufacturerclaimsnocrossreactionobservedwithblood,semen,urine,vaginal secretions,ormenstrualblood

Conclusions
Varioustypesofbiologicalevidenceareencounteredinforensicinvestigationsand severalchallengesmustbetakenintoconsiderationduringprocessingandDNAtesting. ChainofCustodymustbemaintainedduringtheentireprocessofevidencecollection andtestingtoprotectphysicalandlegalintegrity. Serologicalpresumptiveandconfirmatorytestingprovidesinformationofthetypeof biologicalfluidpresentintheevidentiarystain. Numerousmethodsincludingchemicalcolortests,antigenantibodytesting,and microscopicexaminationsareincorporatedinserologytestingofbiologicalevidence stains. Becausesomanydifferenttestsareavailable,specificcrimelaboratoryprotocolsand analystdiscretionshouldbefollowedwhenperformingforensicscreeningandserology. Serologytestingresultsprovidevaluableinformationusedbytheanalysttodetermine whetherDNAorothertestingistobeperformed.

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