Forensic Biology

558 Pages·2015·14.76 MB·English

by  Li, Richard

FORENSIC SCIENCE LI
FORENSIC
BIOLOGY
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S E C O N D E D I T I O N
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E
Over the last several years, new research and developments in analysis methods and practice R
have led to rapid advancements in forensic biology. Identifying critical points of knowledge and
C
E
new methodological approaches in the field, Forensic Biology, Second Edition focuses on foren-
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sic serology and forensic DNA analysis. It provides students and professionals with a scientific N
grounding in biological evidence—both the techniques utilized to identify it and the methodology
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to analyze it.
R I C H A R D L I
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This second edition: C

• Introduces the language of forensic biology, enabling students to become comfortable E
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with usage and terminology
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FORENSIC
• Provides clear explanations of the principles of forensic identification and analysis IO
of biological evidence
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• Explains forensic serology and DNA techniques used in the field and the laboratory IO
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• Discusses the benefits and limitations that apply to various forensic biology techniques G
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• Includes schematic illustrations to clarify concepts Y
BIOLOGY
• Presents three new chapters (Chapters 2, 16, and 17) created for this edition
• Adds more than two hundred new full-color figures
Covering the full scope of forensic biology, the book uses an accessible, straightforward style
designed to enhance students’ education and training so they are prepared, both in the laboratory
and in the field.
S E C O N D E D I T I O N
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New York, NY 10017
an informa business
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w w w . c r c p r e s s . c o m

FORENSIC
BIOLOGY
S E C O N D E D I T I O N
R I C H A R D L I
John Jay College of Criminal Justice
New York, New York, USA
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
CRC Press
Taylor & Francis Group
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Boca Raton, FL 33487-2742
© 2015 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group, an Informa business
No claim to original U.S. Government works
Version Date: 20140818
International Standard Book Number-13: 978-1-4398-8972-5 (eBook - PDF)
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Contents
Preface .......................................................................................................................................xv
Acknowledgments .................................................................................................................xvii
Author ......................................................................................................................................xix
Introduction ............................................................................................................................xxi
SECTION I Biological Evidence
Chapter 1 Crime Scene Investigation of Biological Evidence ...........................................3
1.1 Protection of Crime Scene ........................................................................3
1.2 Recognition of Biological Evidence .........................................................3
1.3 Searches .......................................................................................................7
1.4 Documentation ........................................................................................16
1.5 Chain of Custody .....................................................................................17
1.6 Collection of Biological Evidence ..........................................................18
1.7 Marking Evidence ...................................................................................20
1.8 Packaging and Transportation .............................................................20
1.9 Final Survey and the Release of the Crime Scene ..............................25
1.10 Crime Scene Reconstruction .................................................................29
Bibliography .......................................................................................................30
Chapter 2 Crime Scene Bloodstain Pattern Analysis ......................................................35
2.1 Basic Biological Properties of Human Blood .......................................35
2.2 Formation of Bloodstains .......................................................................35
2.3 Chemical Enhancement and Documentation of Bloodstain
Evidence ....................................................................................................37
2.4 Analyzing Spatter Stains .........................................................................39
2.4.1 Velocity of Blood Droplets .........................................................39
2.4.2 Determining the Directionality of the Stains .........................39
2.4.3 Determining Angles of Impact ................................................40
2.4.4 Determining Area of Origin .....................................................41
2.5 Types of Bloodstain Patterns .................................................................44
2.5.1 Passive Bloodstains ....................................................................44
2.5.2 Transfer Bloodstains .................................................................46
2.5.3 Projected Bloodstains ................................................................46
Bibliography ........................................................................................................51
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Contents
Chapter 3 Forensic Biology: A Subdiscipline of Forensic Science ..................................53
3.1 Common Disciplines of Forensic Laboratory Services ......................53
3.2 Laboratory Analysis of Biological Evidence .........................................53
3.2.1 Identification of Biological Evidence .......................................53
3.2.2 Comparison of Individual Characteristics of
Biological Evidence ....................................................................54
3.2.3 Reporting Results and Expert Testimony ..............................54
3.3 Forensic Science Services Related to Forensic Biology .......................57
3.3.1 Forensic Pathology ....................................................................58
3.3.2 Forensic Anthropology .............................................................58
3.3.3 Forensic Entomology .................................................................60
3.3.4 Forensic Odontology .................................................................64
3.4 Brief History of the Development of Forensic Biology ......................65
3.4.1 Antigen Polymorphism.............................................................65
3.4.2 Protein Polymorphism ..............................................................66
3.4.3 DNA Polymorphism .................................................................66
3.4.3.1 Genes and Related Sequences ...................................66
3.4.3.2 Intergenic Noncoding Sequences .............................66
3.4.3.3 Human DNA Polymorphic Markers .......................68
3.4.3.4 Forensic DNA Polymorphism Profiling .................69
Bibliography ........................................................................................................70
Chapter 4 Sources of Biological Evidence ........................................................................77
4.1 Bodily Fluids ............................................................................................77
4.1.1 Extracellular Nucleic Acids ......................................................77
4.2 Cells ...........................................................................................................81
4.2.1 Cell Surface Markers ..................................................................81
4.2.2 Nucleated Cells ............................................................................81
4.2.3 Mitochondria and Other Organelles ......................................83
4.2.4 Cytosol ........................................................................................84
4.2.4.1 Messenger RNAs ........................................................84
4.2.4.2 MicroRNAs .................................................................85
4.3 Tissues ......................................................................................................87
4.3.1 Skin ..............................................................................................87
4.3.1.1 Biology of Skin ............................................................87
4.3.1.2 Skin as Source of DNA Evidence .............................87
4.3.2 Hair ..............................................................................................88
4.3.2.1 Biology of Hair ...........................................................89
4.3.2.2 Hair as Source of DNA Evidence ..............................91
4.3.3 Bone .............................................................................................92
4.3.3.1 Biology of Bone ...........................................................92
4.3.3.2 Bone as Source of DNA Evidence ............................94
4.3.4 Teeth ............................................................................................98
4.3.4.1 Biology of Teeth ..........................................................98
4.3.4.2 Teeth as Source of DNA Evidence .........................100
Bibliography ......................................................................................................102
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Contents
SECTION II Basic Techniques in Forensic Biology
Chapter 5 Nucleic Acid Extraction ...................................................................................111
5.1 Basic Principles of DNA Extraction ....................................................111
5.1.1 Cell and Tissue Disruption ......................................................111
5.1.2 Lysis of Cellular and Organelle Membranes .........................114
5.1.3 Removal of Proteins and Cytoplasmic Constituents ...........114
5.1.4 Storage of DNA Solutions ........................................................115
5.1.5 Contamination ..........................................................................115
5.2 Methods of DNA Extraction ................................................................115
5.2.1 Extraction with Phenol–Chloroform .....................................115
5.2.1.1 Cell Lysis and Protein Digestion .............................115
5.2.1.2 Extraction with Organic Solvents ...........................115
5.2.1.3 Concentrating DNA ..................................................115
5.2.2 Extraction by Boiling Lysis and Chelation ............................117
5.2.2.1 Washing ......................................................................117
5.2.2.2 Boiling .........................................................................117
5.2.2.3 Centrifugation ...........................................................117
5.2.3 Silica-Based Extraction ............................................................118
5.2.3.1 Cell Lysis and Protein Digestion .............................118
5.2.3.2 DNA Adsorption onto Silica ...................................118
5.2.3.3 Washing ......................................................................119
5.2.3.4 Elution of DNA ..........................................................119
5.2.4 Differential Extraction .............................................................121
5.3 Essential Features of RNA ...................................................................122
5.4 Methods of RNA Extraction ...............................................................124
5.4.1 RNA–DNA Coextraction .......................................................124
5.4.2 miRNA Extraction ..................................................................124
Bibliography .....................................................................................................127
Chapter 6 DNA Quantitation ............................................................................................133
6.1 Slot Blot Assay ........................................................................................133
6.2 Fluorescent Intercalating Dye Assay ..................................................134
6.3 Quantitative PCR Assay ........................................................................137
6.3.1 Real-Time Quantitative PCR...................................................137
6.3.1.1 TaqMan Method .......................................................138
Bibliography ......................................................................................................140
Chapter 7 Amplification by Polymerase Chain Reaction .............................................143
7.1 Denaturation and Renaturation of DNA ............................................143
7.2 Basic Principles of Polymerase Chain Reaction ................................143
7.3 Essential PCR Components ..................................................................145
7.3.1 Thermostable DNA Polymerases ............................................145
7.3.2 PCR Primers ..............................................................................146
7.3.3 Other Components ...................................................................147
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Contents
7.4 Cycle Parameters ....................................................................................147
7.5 Factors Affecting PCR ...........................................................................149
7.5.1 Template Quality ......................................................................149
7.5.2 Inhibitors ...................................................................................149
7.5.3 Contamination ..........................................................................149
7.6 Reverse Transcriptase PCR for RNA-Based Assays .........................150
7.6.1 Reverse Transcription .............................................................150
7.6.2 Oligodeoxynucleotide Priming ..............................................151
7.6.3 Reverse Transcriptase PCR .....................................................151
Bibliography ......................................................................................................153
Chapter 8 DNA Electrophoresis .......................................................................................159
8.1 Basic Principles ......................................................................................159
8.2 Supporting Matrices ..............................................................................159
8.2.1 Agarose .......................................................................................160
8.2.2 Polyacrylamide ..........................................................................161
8.2.2.1 Polymerization Reaction ..........................................161
8.2.2.2 Cross-Linking Reaction ...........................................161
8.2.2.3 Denaturing Polyacrylamide Electrophoresis ........163
8.3 Apparatus and Forensic Applications .................................................163
8.3.1 Slab Gel Electrophoresis ..........................................................163
8.3.1.1 Agarose Gel Electrophoresis ....................................163
8.3.1.2 Polyacrylamide Gel Electrophoresis ......................164
8.3.2 Capillary Electrophoresis .......................................................164
8.3.3 Microfluidic Devices ................................................................165
8.3.3.1 Modular Microfluidic Devices ................................167
8.3.3.2 Integrated Microfluidic Devices..............................168
8.4 Estimation of DNA Size ........................................................................168
8.4.1 Relative Mobility .......................................................................168
8.4.2 Local Southern Method ...........................................................170
Bibliography ......................................................................................................171
Chapter 9 Detection Methods ...........................................................................................175
9.1 Direct Detection of DNA in Gels ........................................................175
9.1.1 Fluorescent Intercalating Dye Staining .................................175
9.1.2 Silver Staining ...........................................................................175
9.2 Detection of DNA Probes in Hybridization-Based Assays ..............177
9.2.1 Radioisotope Labeled Probes ..................................................177
9.2.2 Enzyme-Conjugated Probe with
Chemiluminescence Reporting System ................................180
9.2.3 Biotinylation of DNA with Colorimetric
Reporting Systems ...................................................................180
9.2.3.1 Biotin ..........................................................................180
9.2.3.2 Enzyme-Conjugated Avidin ...................................180
9.2.3.3 Reporter Enzyme Assay ...........................................181
9.3 Detection Methods for PCR-Based Assays ........................................182
9.3.1 Fluorescence Labeling ..............................................................182
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9.3.1.1 Fluorescent Dyes .......................................................182
9.3.1.2 Labeling Methods......................................................182
9.3.1.3 Fluorophore Detection .............................................185
Bibliography ......................................................................................................186
Chapter 10 Serology Concepts ............................................................................................189
10.1 Serological Reagents ..............................................................................189
10.1.1 Immunogens and Antigens .....................................................189
10.1.2 Antibodies..................................................................................189
10.1.2.1 Polyclonal Antibodies ...............................................191
10.1.2.2 Monoclonal Antibodies ............................................191
10.1.2.3 Antiglobulins ............................................................192
10.2 Strength of Antigen–Antibody Binding .............................................193
10.3 Antigen–Antibody Binding Reactions ................................................193
10.3.1 Primary Reactions ...................................................................194
10.3.2 Secondary Reactions ...............................................................194
10.3.2.1 Precipitation ...............................................................195
10.3.2.2 Agglutination ............................................................197
Bibliography .....................................................................................................198
Chapter 11 Serology Techniques: Past, Current, and Future .........................................199
11.1 Introduction to Forensic Serology ......................................................199
11.1.1 The Scope of Forensic Serology ..............................................199
11.1.2 Class Characteristics and Individual Characteristics
of Biological Evidence .............................................................199
11.1.3 Presumptive and Confirmatory Assays ................................200
11.1.4 Primary and Secondary Binding Assays ..............................200
11.2 Primary Binding Assays ......................................................................201
11.2.1 Enzyme-Linked Immunosorbent Assay (ELISA) ...............201
11.2.2 Immunochromatographic Assays .........................................202
11.3 Secondary Binding Assays ...................................................................204
11.3.1 Precipitation-Based Assays .....................................................204
11.3.1.1 Immunodiffusion .....................................................204
11.3.1.2 Immunoelectrophoretic Methods ..........................206
11.3.2 Agglutination-Based Assays ...................................................208
11.4 DNA Methylation Assays for Bodily Fluid Identification ................210
11.5 Forensic Applications of RNA-Based Assays and RNA Profiling.....212
11.5.1 Messenger RNA-Based Assays ................................................212
11.5.2 MicroRNA-Based Assays .........................................................214
11.6 Proteomic Approaches Using Mass Spectrometry for Bodily
Fluid Identification ................................................................................215
11.6.1 Mass Spectrometric Instrumentation for
Protein Analysis .....................................................................215
11.6.2 Analysis Strategies for Protein Identification .......................216
11.7 Microbial DNA Analysis for Bodily Fluid Identification .................217
11.8 Nondestructive Assays for the Identification of Bodily Fluids .......220
Bibliography ......................................................................................................221
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Contents
SECTION III Identification of Biological Evidence
Chapter 12 Identification of Blood .....................................................................................231
12.1 Biological Properties .............................................................................231
12.1.1 Red Blood Cells .........................................................................231
12.1.2 White Blood Cells .....................................................................231
12.1.3 Platelets .......................................................................................231
12.2 Presumptive Assays for Identification ...............................................232
12.2.1 Mechanisms of Presumptive Assays .....................................232
12.2.1.1 Oxidation–Reduction Reactions ............................234
12.2.2 Colorimetric Assays ................................................................234
12.2.2.1 Phenolphthalin Assay ..............................................234
12.2.2.2 Leucomalachite Green (LMG) Assay ....................234
12.2.2.3 Benzidine and Derivatives ......................................234
12.2.3 Chemiluminescence and Fluorescence Assays ....................236
12.2.3.1 Luminol (3-Aminophthalhydrazide).....................237
12.2.3.2 Fluorescin ..................................................................238
12.2.4 Factors Affecting Presumptive Assay Results ......................238
12.2.4.1 Oxidants ....................................................................238
12.2.4.2 Plant Peroxidases ......................................................239
12.2.4.3 Reductants .................................................................239
12.3 Confirmatory Assays for Identification .............................................239
12.3.1 Microcrystal Assays.................................................................239
12.3.1.1 Hemochromagen Crystal Assay .............................239
12.3.1.2 Hematin Crystal Assay ............................................240
12.3.2 Other Assays .............................................................................240
Bibliography .....................................................................................................241
Chapter 13 Species Identification ......................................................................................245
13.1 General Considerations .......................................................................245
13.1.1 Types of Antibodies .................................................................245
13.1.2 Titration of Antibodies ...........................................................246
13.1.3 Antibody Specificity ................................................................246
13.1.4 Optimal Conditions for Antigen–Antibody Binding .........247
13.2 Assays .....................................................................................................247
13.2.1 Immunochromatographic Assays .........................................247
13.2.1.1 Identification of Human
Hemoglobin Protein ........................................247
13.2.1.2 Identification of Human Glycophorin
A Protein ................................................................248
13.2.2 Double Immunodiffusion Assays ...........................................251
13.2.2.1 Ring Assay ..................................................................251
13.2.2.2 Ouchterlony Assay ....................................................251
13.2.3 Crossed-Over Electrophoresis ................................................251
Bibliography .....................................................................................................255
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