Molecular Diagnostics - Technologies, Markets and Companies

Table of Contents

Part I

0. Executive Summary 21

1. Introduction 23

• Definitions and scope of the subject23
• Historical evolution of molecular diagnostics23
• Molecular biology relevant to molecular diagnostics24
• Genome24
• DNA24
• DNA polymerases25
• Restriction endonucleases25
• DNA methylation25
• RNA26
• RNA polymerases26
• MicroRNAs27
• DNA transcription27
• Chromosomes27
• Telomeres28
• Mitochondrial DNA28
• Genes29
• The genetic code29
• Gene expression29
• DNA sequences30
• Junk DNA30
• Single nucleotide polymorphisms30
• Genotype and haplotypes31
• Replication of the DNA helix31
• Proteins32
• Proteomics32
• Monoclonal antibodies32
• Aptamers32
• Basics of molecular diagnostics33
• Tracking DNA: the Southern blot33
• Pulsed-field gel electrophoresis34
• DNA Probes34
• The polymerase chain reaction34
• Basic Principles of PCR34
• Target selection35
• Detection of amplified DNA35
• Impact of human genome project on molecular diagnostics36
• Genetic variations in the human genome36
• Insertions and deletions in the human genome36
• Large scale variation in human genome37
• Variation in copy number in the human genome37
• Structural variations in the human genome38
• Mapping and sequencing of structural variation from human genomes39
• 1000 Genomes Project40
• Human Variome Project40
• Systems biology approach to molecular diagnostics41
• Biomarkers42
• Applications of molecular diagnostics42

2. Molecular Diagnostic Technologies 45

• Introduction45
• DNA extraction45
• Transrenal DNA45
• Sample preparation46
• Pressure Cycling Technology46
• Membrane immobilization of nucleic acids46
• Automation of sample preparation in molecular diagnostics47
• ABI PRISM 6700 Automated Nucleic Acid Workstation47
• BioRobot technology47
• COBAS AmpliPrep System47
• GENESIS FE500 Workcell47
• GeneMole48
• PCR BioCube48
• QIAsymphony48
• Tigris instrument system48
• Techniques for sample preparation that are suitable for automation49
• Xtra Amp Genomic DNA Extraction49
• Extraction of DNA from paraffin sections49
• Dynabead technology49
• Pressure Cycling Technology50
• SamPrep50
• Use of magnetic particles for automation in genome analysis50
• Companies involved in nucleic acid isolation51
• Novel PCR methods52
• Addressing limitations of PCR52
• Real-time PCR systems52
• Dyes used in real-time PCR53
• Commercially available real-time PCR systems54
• LightCycler PCR system54
• LightUp probes based on real-time PCR55
• Applications of real-time PCR55
• Limitations of real-time PCR56
• Improving the reliability of low level DNA analysis by real-time PCR56
• Guidelines for real-time quantitative PCR56
• Future applications of real-time Q-PCR57
• Reverse transcriptase (RT)-PCR57
• Standardized reverse transcriptase PCR58
• Single cell PCR58
• LATE-PCR58
• Cold-PCR59
• AmpliGrid-System59
• Digital PCR59
• Long and accurate PCR60
• Combined PCR-ELISA60
• Monitoring of gene amplification in molecular diagnostics61
• Non-PCR nucleic acid amplification methods61
• Linked Linear Amplification61
• Multiplex Ligation-Dependent Probe Amplification61
• Transcription mediated amplification62
• Rapid analysis of gene expression62
• WAVE nucleic acid fragment analysis system62
• DNA probes with conjugated minor groove binder63
• Rolling circle amplification technology63
• Gene-based diagnostics through RCAT65
• RCAT-immunodiagnostics65
• RCAT-biochips65
• RCAT-pharmacogenomics66
• Circle-to-circle amplification66
• Ramification amplification method66
• Single Primer Isothermal Amplification66
• Isothermal reaction for amplification of oligonucleotides67
• ICAN (Isothermal and Chimeric primer-initiated Amplification of Nucleic Acids)67
• Technologies for signal amplification68
• 3 DNA dendrimer signal amplification68
• Hybridization signal amplification method69
• Signal mediated amplification of RNA technology70
• Invader assays70
• Hybrid Capture technology71
• Branched DNA test72
• Tyramide signal amplification73
• Non-enzymatic signal amplification technologies73
• Direct molecular analysis without amplification74
• Direct detection of dsDNA75
• Multiplex assays75
• Fluorescent in situ hybridization75
• Modifications of FISH77
• Direct visual in situ hybridization77
• Direct labeled Satellite FISH probes78
• Comparative genomic hybridization78
• Primed in situ labeling78
• Interphase FISH79
• FISH with telomere-specific probes79
• Multicolor FISH79
• Automation of FISH80
• Companies involved in FISH diagnostics80
• RNA diagnostics81
• Branched-chain DNA assay for measurement of RNA82
• Cycling probe technology83
• Invader RNA assays83
• Linear RNA amplification83
• Non-isotopic RNase cleavage assay84
• Nucleic acid sequence-based amplification84
• Q Beta replicase system86
• Solid Phase Transcription Chain Reaction86
• Transcriptome analysis86
• Visualization of mRNA expression in vivo87
• MicroRNA diagnostics87
• Real-time PCR for expression profiling of miRNAs87
• Microarray vs quantitative PCR foro measuring miRNAs88
• Use of LNA to explore miRNA88
• Nuclease Protection Assay to measure miRNA expression89
• Microarrays for analysis of miRNA gene expression89
• Modification of in situ hybridization for detection of miRNAs90
• Whole genome amplification90
• Companies that provide technologies for whole genome amplification90
• QIAGEN' s Repli-G system91
• GenomePlex™ Whole Genome Amplification91
• DNA sequencing92
• Companies involved in sequencing93
• Applications of next generation sequencing in molecular diagnostics94
• Genome-wide approach for chromatin mapping94
• Mitochondrial sequencing94
• Identification of unknown DNA sequences95
• Optical mapping95
• Gene expression analysis96
• Gene expression profiling on whole blood samples96
• Gene expression patterns of white blood cells97
• Gene expression profiling based on alternative RNA splicing97
• MAUI (MicroArray User Interface) hybridization98
• Monitoring in vivo gene expression by molecular imaging98
• Serial analysis of gene expression (SAGE)98
• Single-cell gene expression analysis98
• T cell receptor expression analysis99
• Tangerine"! expression profiling99
• Whole genome expression array100
• Ziplex"! system100
• Companies involved in gene expression analysis101
• Peptide nucleic acid technology102
• Use of PNA with fluorescence in situ hybridization102
• PNA and PCR103
• Use of PNA with biosensors103
• PNA-based PD-loop technology104
• PNA-DNA hybrid quadruplexes104
• Companies Involved in PNA Diagnostics104
• Locked nucleic acids105
• Electrochemical detection of DNA105
• Mediated nucleic acid oxidation106
• Detection of hybridized nucleic acid with cyclic voltametry106
• Electrochemical detection based on Toshiba' s CMOS technology107
• Concluding remarks on electrochemical DNA detection107
• Scorpions"! technology107
• The Scorpions reaction107
• Applications of Scorpion108

3. Biochips, Biosensors, and Molecular Labels 109

• Introduction to biochip technology109
• Applications of biochips in diagnostics109
• GeneChip110
• GeneChip Human Genome Arrays111
• AmpliChip CYP450111
• Electronic detection of nucleic acids on microarrays111
• Microchip capillary electrophoresis112
• Strand displacement amplification on a biochip112
• Rolling circle amplification on DNA microarrays112
• Fast PCR biochip112
• Multiplex microarray-enhanced PCR for DNA analysis113
• Multiplexed Molecular Profiling113
• Universal DNA microarray combining PCR and ligase detection reaction114
• Genomewide association scans114
• Whole genome chips/microarrays114
• Transposon insertion site profiling chip115
• Standardizing the microarrays115
• Companies involved in developing biochip technology for diagnostics116
• Future of biochip technology for molecular diagnostics117
• Microfluidic chips117
• Fish-on-chip118
• Lab-on-a-chip118
• LabCD118
• Micronics' microfluidic technology119
• Microfluidic automated DNA analysis using PCR119
• Microfluidic chips integrated with PET119
• Companies developing microfluidic technologies120
• Biosensor technologies120
• Classification of biosensor technologies121
• DNA-based biosensors122
• DNA hybridization biosensor chips122
• PCR-free DNA biosensor122
• DNA based biosensor to detects metallic ions123
• Genetically engineered B lymphocytes123
• Biosensors immunoassays123
• PNA (peptide nucleic acid)-based biosensors124
• Protein-based biosensors124
• Antibody biosensors124
• Cell-based biosensors (cytosensors)124
• Multicell biosensors125
• Microbial biosensors125
• Optical biosensors126
• Surface plasmon resonance technology126
• Label-free optical biosensor127
• Microsensors using with nano/microelectronic communications technology127
• Electrochemical sensors127
• Enzyme electrodes for biosensing127
• Conductometric sensors128
• Electrochemical genosensors128
• Electrochemical nanobiosensor128
• Bioelectronic sensors129
• Phototransistor biochip biosensor129
• Ribozyme-based sensors129
• RiboReporters130
• Concluding remarks and future prospects of biosensor technology130
• Companies developing biosensors for molecular diagnostics131
• Molecular labels and detection132
• Detection technologies for molecular labels133
• Fluorescence and chemiluminescence133
• Fluorescence technologies for label detection134
• Companies with fluorescence and chemiluminescence products134
• Molecular beacons135
• The Green fluorescent protein136
• Multiophoton detection radioimmunoassay137
• Multi-pixel photon counter137
• Enzyme labels and detection by fluorescence137
• Phase-sensitive flow cytometry138
• Microtransponder-based DNA diagnostics138
• Laboratory Multiple Analyte Profile139
• Multiple labels139
• Protein-DNA chimeras for detection of small numbers of molecules140
• Single molecule detection140
• Atomic force microscopy141
• Capillary electrophoresis141
• Confocal laser scanning141
• Spectrally resolved fluorescence lifetime imaging microscopy141
• Molecular imaging142
• Basic research in molecular imaging142
• Devices for molecular imaging143
• Molecular imaging in clinical practice143
• Challenges and future prospects of molecular imaging143
• Companies involved in molecular imaging143
• Nanobiotechnology for molecular diagnostics144
• Magnetic nanoparticles145
• Gold nanoparticles146
• Quantum dot technology146
• Nanotechnology on a chip147
• Nanogen' s NanoChip148
• Fullerene photodetectors for chemiluminescence detection on microfluidic chip148
• Diagnostics based on nanopore technology148
• Nanosensors148
• Quartz nanobalance biosensor149
• PEBBLE nanosensors149
• Nanosensors for glucose monitoring149
• Cantilever arrays150
• Resonance Light Scattering technology150
• DNA nanomachines for molecular diagnostics151
• Nanobarcodes technology for molecular diagnostics151
• Qdot nanobarcode for multiplexed gene expression profiling152
• Role of nanobiotechnology in improving molecular diagnostics152
• Companies involved in nanomolecular diagnostics152
• Concluding remarks about nanodiagnostics155
• Future prospects of nanodiagnostics155

4. Proteomic Technologies for Molecular Diagnostics 157

• Introduction157
• Proteomic technologies157
• Biomarkers of disease157
• Proteomic tools for biomarkers157
• Search for biomarkers in body fluids158
• Captamers with proximity extension assay for proteins158
• Cyclical amplification of proteins158
• Detection of misfolded proteins by ELISA with exponential signal amplification159
• Diagnostics based on designed repeat proteins159
• Differential Peptide Display159
• Light-switching excimer probes160
• MALDI-TOF Mass Spectrometry160
• Molecular beacon aptamer161
• Molecular beacon assay161
• Proteomic patterns161
• Real-time PCR for protein quantification163
• Protein biochip technologies163
• ProteinChip164
• LabChip for protein analysis165
• TRINECTIN proteome chip165
• Protein chips for antigen-antibody interactions molecular diagnostics165
• Microfluidic devices for proteomics-based diagnostics166
• Nanotechnology-based protein biochips/microarrays166
• Nanoparticle protein chip166
• Protein nanobiochip166
• Protein biochips based on fluorescence planar wave guide technology167
• New developments in protein chips/microarrays167
• Antibody microarrays168
• Aptamer-based protein biochip168
• Multiplexed Protein Profiling on Microarrays168
• Proteomic pattern analysis169
• Single molecule array169
• Viral protein chip169
• Commercial development of protein chips for molecular diagnostics170
• Proteome Identification Kit171
• Laser capture microdissection (LCM)171
• LCM technology171
• Applications of LCM in molecular diagnostics172
• Proteomic diagnosis of CNS disorders172
• Cerebrospinal fluids tests based on proteomics172
• Urine tests for CNS disorders based on proteins in urine173
• Diagnosis of CNS disorders by examination of proteins in the blood173
• Diagnosis of CNS disorders by examination of proteins in tears174
• Role of proteomics in the diagnosis of Alzheimer' s disease175
• Role of proteomics in the diagnosis of Creutzfeldt-Jakob disease175
• Future prospects of use of proteomics for diagnosis of CNS disorders175
• Concluding remarks on the use of proteomics in diagnostics175

5. Molecular Diagnosis of Genetic Disorders 177

• Introduction177
• Cytogenetics178
• FISH with probes to the telomeres178
• Single copy FISH probes178
• Comparative genomic hybridization179
• Use of biochips in genetic disorders179
• Representational oligonucleotide microarray analysis180
• SignatureChipR-based diagnostics for cytogenetic abnormalities180
• Diagnosis of genomic rearrangements by multiplex PCR180
• Quantitative fluorescent PCR180
• Mutation detection technologies181
• PCR-based methods for mutation detection182
• Cleavase Fragment Length Polymorphism182
• Direct dideoxy DNA sequencing182
• Digital Genetic Analysis (DGA)182
• Fluorescence-based directed termination PCR183
• Heteroduplex analysis183
• Restriction fragment length polymorphism184
• Single-stranded conformation polymorphism (SSCP) analysis184
• TaqMan real-time PCR184
• Non-PCR methods for mutation detection185
• Arrayed primer extension185
• BEAMing (beads, emulsion, amplification, and magnetics)185
• ELISA-protein truncation test186
• Enzymatic mutation detection186
• Specific anchor nucleotide incorporation186
• Conversion analysis for mutation detection187
• Biochip technologies for mutation detection187
• Combination of FISH and gene chips187
• Haplotype Specific Extraction188
• Technologies for SNP analysis188
• DNA sequencing189
• Electrochemical DNA probes189
• Use of NanoChip for detection of SNPs190
• Single base extension-tag array190
• Laboratory Multiple Analyte Profile190
• SNP genotyping with gold nanoparticle probes191
• PCR-CTPP (confronting two-pair primers)191
• Peptide nucleic acid probes for SNP detection191
• SNP genotyping on a genome-wide amplified DOP-PCR template192
• Pyrosequencing192
• Reversed enzyme activity DNA interrogation test192
• Smart amplification process version 2193
• Zinc finger proteins193
• UCAN method (Takara Biomedical)194
• Biochip and microarray-based detection of SNPs194
• SNP genotyping by MassARRAY194
• Electronic dot blot assay194
• Biochip combining BeadArray and ZipCode technologies195
• SNP-IT primer-extension technology195
• OmniScan SNP genotyping196
• Affymetrix SNP genotyping array196
• Concluding remarks on SNP genotyping196
• Limitations of SNP in genetic testing196
• Haplotyping versus SNP genotyping197
• Determination of copy number variations197
• Companies involved in developing technologies/products for SNP analysis198
• Prenatal DNA diagnosis200
• Amniocentesis200
• Chorionic villus sampling200
• Separating fetal cells in maternal blood for genetic diagnosis200
• Antenatal screening for Down' s syndrome201
• Fetal DNA in maternal blood201
• Molecular methods for prenatal diagnosis202
• aCGH for prenatal diagnosis202
• BAC HD Scan test202
• FISH for prenatal diagnosis203
• PCR for prenatal diagnosis203
• Plasma DNA sequencing to detect fetal chromosomal aneuploidies203
• In vivo gene expression analysis of the living human fetus204
• Noninvasive prenatal diagnosis of monogenic diseases204
• Digital relative mutation dosage204
• Massively parallel plasma DNA sequencing204
• Applications of prenatal diagnosis205
• Diagnosis of congenital infections206
• Diagnosis of eclampsia206
• Use of transrenal DNA for prenatal testing206
• Preimplantation genetic diagnosis207
• Technologies for preimplantation genetic diagnosis (PGD)207
• PCR for preimplantation genetic diagnosis207
• FISH for preimplantation genetic diagnosis207
• Microarrays for preimplantation genetic diagnosis208
• Conditions detected by preimplantation genetic diagnosis208
• The future of preimplantation genetic diagnosis209
• Companies involved in prenatal/preimplantation diagnosis210
• Cystic fibrosis210
• Detection of CFTR gene mutations211
• CFTR technologies of various companies211
• Genzyme' s CF gene sequencing212
• CF Plus™ Tag-It Cystic Fibrosis Kit212
• Asuragen' s bead array test213
• The Ambry CF Test213
• Biochip for CF diagnosis213
• Identification of CF variants by PCR/Oligonucleotide Ligation Assay214
• MassARRAYR system for high-throughput CFTR testing214
• Serum proteomic signature for CF using antibody microarrays214
• Guidelines for genetic screening for CF214
• Congenital adrenal hyperplasia215
• Primary immunodeficiencies215
• Hematological disorders216
• Hemoglobinopathies216
• Sickle cell anemia216
• Thalassemia217
• Paroxysmal nocturnal hemoglobinuria217
• Hemophilia218
• Hereditary hemochromatosis218
• Polycystic kidney disease218
• Hereditary metabolic disorders219
• Lesch-Nyhan Syndrome219
• Gaucher' s Disease219
• Acute Intermittent Porphyria220
• Phenylketonuria220
• Hereditary periodic fever220
• Achondroplasia220
• Molecular diagnosis of cardiovascular disorders221
• Coronary Heart Disease222
• Cardiomyopathy222
• Familial Hypertrophic Cardiomyopathy223
• Idiopathic dilated cardiomyopathy223
• Cardiac Arrhythmias223
• Long Q-T Syndrome223
• Familial atrial fibrillation224
• Idiopathic ventricular fibrillation224
• Congestive heart failure224
• Hypertension224
• Disturbances of blood lipids225
• Familial dyslipoproteinemias225
• Hypercholesterolemia225
• Thrombotic disorders226
• Factor V Leiden mutation226
• Pulmonary embolism226
• Molecular diagnosis of eye diseases227
• Molecular diagnosis of retinitis pigmentosa227
• Genetic screening for glaucoma227
• Role of molecular diagnostics in rheumatoid arthritis228
• Molecular diagnosis of neurogenetic disorders228
• Alzheimer' s disease230
• Down syndrome230
• Parkinson' s disease231
• Spinal muscular atrophy232
• Duchenne and Becker muscular dystrophy232
• Triple repeat disorders233
• Huntington disease233
• Fragile X syndrome234
• Charcot-Marie Tooth disease234
• Hereditary neuropathy with liability to pressure palsies235
• eNOS gene polymorphisms as predictor of cerebral aneurysm rupture235
• Mitochondrial disorders affecting the nervous system235
• Genetic testing for disease predisposition236
• Direct-to-consumer genetic tests236

6. Molecular Diagnosis of Infections 238

• Introduction238
• Molecular techniques for the diagnosis of infections238
• Antibody-enhanced microplate hybridization assays239
• Biosensors for detection of microorganisms239
• Ibis T5000"! Biosensor System239
• DNA enzyme immunoassay240
• DNA biochip/microarray in diagnosis of infections240
• DNA-based typing methods241
• Restriction fragment length polymorphism analysis241
• Ribotyping241
• Random amplified polymorphic DNA241
• Combinatorial DNA melting assay241
• Electrochemical detection of pathogens242
• Ligase chain reaction242
• Mass spectrometry for microbial identification242
• Metagenomic pyrosequencing242
• Multiplex PCR for detection of infections244
• LightCyclerR SeptiFast Test244
• VYOOR Sepsis Test244
• Dual priming oligonucleotide for multiplex PCR245
• NASBA for detection of microorganisms245
• Nucleic acid probes245
• Neutrophil CD11b expression as a diagnostic marker246
• Optical Mapping246
• PNA-FISH for diagnosis of infections246
• Quantitative reverse-transcription PCR for bacterial diagnostics247
• Rupture event scanning247
• Real-time single-molecule imaging of virus particles247
• Single-strand conformational polymorphism247
• SmartGene platform for identifying pathogens based on genetic sequences248
• Tessera array technology248
• Applications, advantages and limitations of molecular diagnostics248
• Molecular diagnostics versus other microbial detection technologies248
• Advantages of nucleic acid-based diagnostics in infections249
• Drawbacks of nucleic acid-based diagnostics in infections249
• Nanotechnology for detection of infectious agents250
• Bacterial and fungal infections250
• Mycobacterium tuberculosis252
• Conventional diagnosis of tuberculosis252
• Microscopic Observation Drug Susceptible Assay for tuberculosis252
• Molecular diagnostics for tuberculosis253
• Combined tuberculin testing and ELISpotPLUS assay254
• Biomarkers for tuberculosis255
• Diagnosis of drug-resistant M. tuberculosis infection255
• Cost-effectiveness of PCR in tuberculosis screening256
• Other mycobacteria256
• Chlamydial infections256
• Neisseria gonorrhoeae258
• Bacteria associated with bacterial vaginosis258
• Streptococcal infections258
• Group B Streptococci258
• Streptococcus pyogenes and Streptococcus dysgalactiae259
• Pseudomonas aeruginosa259
• Helicobacter pylori260
• Lyme disease260
• Mycoplasmas261
• Fungal infections261
• Viral infections262
• HIV/AIDS263
• Diagnosis of HIV263
• Neonatal screening of infants of HIV-positive mothers264
• Screening of cadaveric tissue donors264
• Detection of HIV provirus264
• Resolution of indeterminate Western blot264
• Global Surveillance of HIV-1 genetic variations265
• Genotyping for drug-resistance in HIV265
• Phenotyping as predictor of drug susceptibility/resistance in HIV266
• Tests used for quantification of HIV267
• Conclusions about HIV genotyping267
• Hepatitis viruses268
• Hepatitis A virus268
• Hepatitis B virus269
• Hepatitis C virus269
• Detection and quantification of HCV RNA270
• Quantification of HCV RNA levels as a guide to antiviral therapy271
• Electrochemical DNA chip for diagnosis of HCV271
• HCV Genotyping as a guide to therapy271
• Enteroviruses272
• Adenoviruses273
• Rhinoviruses273
• Herpes viruses273
• Herpes simplex virus273
• Genital and neonatal herpes simplex274
• Human cytomegalovirus infections274
• Epstein-Barr virus274
• Human papilloma virus275
• Molecular diagnostics for HPV275
• Detection of encephalitis viruses276
• West Nile and St. Louis encephalitis276
• Venezuelan equine encephalitis virus276
• Protozoal infections277
• Amebiasis277
• Cryptosporidium parvum277
• Malaria277
• Neurocysticercosis278
• Pneumocystis carinii278
• Toxoplasmosis278
• Infections of various systems279
• CNS infections279
• Molecular diagnosis in bacterial meningitis279
• Molecular diagnosis in herpes simplex encephalitis279
• Diagnosis of transmissible spongiform encephalopathies280
• Molecular diagnosis of respiratory viruses281
• SARS-associated coronavirus281
• Influenza viruses282
• Avian influenza284
• H1N1 influenza287
• Gastrointestinal infections289
• Periodontal infections290
• Diagnosis of urinary infections by a biosensor291
• Role of molecular diagnostics in septicemia291
• Limitations and needs of diagnostics for infections292
• Differentiation between live and antibiotic-killed bacteria292
• Cell-based methods for identifying pathogenic microorganisms293
• Cell-based virus assays293
• Cell-based detection of host response to infection293
• Role of molecular diagnostics in hospital acquired infections293
• Detection of hospital-acquired bacterial infections294
• Detection of methicillin-resistant S. aureus294
• Detection of vancomycin-resistant enterococci295
• Detection of hospital-acquired C. difficile295
• Bacterial genome sequencing in antimicrobial resistance295
• Detection of hospital-acquired viral infections296
• Molecular diagnosis of BK virus296
• Diagnosis of hospital-acquired rotavirus gastroenteritis296
• Molecular diagnostics and the microbiome297
• Human Microbiome Project297
• Application of metagenomics to study of the microbiome297
• MicroBiome Analysis Center298
• Concluding remarks and future prospects of diagnosis of infections298
• Rapid point-of-care diagnosis of infection299
• Diagnosis of viruses using protein fingerprinting301
• QIAplex PCR multiplex technology301
• Companies involved in molecular diagnosis of infectious diseases301

7. Molecular Diagnosis of Cancer 304

• Introduction304
• Cancer Genetics304
• Oncogenes304
• Tumor Suppressor Genes305
• p53306
• p16306
• Viruses and cancer307
• Detecting viral agents in cancer307
• Conventional cancer diagnosis308
• Molecular techniques for cancer diagnosis309
• Genome analysis at the molecular level310
• Mutation detection at molecular level311
• Expression profiling of tumor cells sorted by flow cytometry311
• MicroRNA expression profiling to classify human cancers311
• Biomarkers in cancer312
• Circulating nucleosomes in serum of cancer patients312
• Detection of DNA methylation313
• eTag assay system for cancer biomarkers315
• HAAH as a biomarker for cancer315
• LigAmp for detection of gene mutations in cancer315
• Mitochondrial DNA as a cancer biomarker316
• Oncoproteins as biomarkers for cancer316
• Sequencing-based approaches for detection of cancer biomarkers317
• Molecular fingerprinting of cancer317
• Fluorescent in situ hybridization318
• Genetic analysis of cancer318
• Comparative genomic hybridization in cancer diagnostics318
• Loss of heterozygosity319
• Digital karyotyping319
• Gene expression profiles predict chromosomal instability in tumors319
• PCR Techniques320
• Realtime quantitative PCR for diagnosis of cancer320
• Cold-PCR320
• Antibody-based diagnosis of cancer321
• Monoclonal antibodies for diagnosis of cancer321
• Recombinant antibodies as a novel approach to cancer diagnosis321
• Combined immunological and nucleic acid tests322
• Combination of MAbs and RT-PCR322
• Immunobead RT-PCR322
• Assays for determining susceptibility to cancer322
• Gene expression profiling in cancer322
• Microarrays for gene expression profiling in cancer323
• Serial analysis of gene expression (SAGE)323
• DNA tags for finding genes expressed in cancer324
• Suppression subtractive hybridization324
• Measurement of telomerase activity324
• Detection of cancer cells in blood of patients with solid tumors325
• Epithelial aggregate separation and isolation326
• Proteomic technologies for the molecular diagnosis of cancer327
• Proteomic technologies for tumor biomarkers327
• Affibodies as contrast agents for imaging in cancer327
• Aptamer-based technology for protein signatures of cancer cells328
• Aptamers for combined diagnosis and therapeutics of cancer328
• Automated image analysis of nuclear protein distribution329
• Laser capture microdissection in oncology329
• Layered expression scanning330
• Survivin and molecular diagnosis of cancer330
• Biochip/microarrays for cancer diagnosis330
• Role of DNA microarrays in gene expression profiling331
• Biochip detection of FHIT gene331
• Nanobiotechnology for early detection of cancer332
• Detection of nanoparticle self assembly in tumors by MRI332
• Differentiation between normal and cancer cells by nanosensors332
• Magnetic nanoparticle probes332
• Quantum dots for early detection of cancer333
• Molecular imaging of cancer333
• In vivo tumor illumination by adenoviral-GFP334
• PET for in vivo molecular diagnosis of cancer334
• Xenon-enhanced MRI334
• Optical systems for in vivo molecular imaging of cancer334
• Detection of micrometastases335
• Molecular diagnosis of cancers of various organs335
• Brain tumors336
• Molecular diagnostic methods for brain tumors336
• Glioblastoma multiforme336
• Circulating microvesicles as biomarkers of glioblastoma337
• Combination of neuroimaging and DNA microarray analysis in GBM337
• Medulloblastoma338
• Oligodendroglioma338
• Advantages and limitations of molecular diagnosis of brain tumors338
• Breast cancer338
• Breast cancer genes339
• Molecular diagnostic tests for breast cancer340
• Mouse ESC-based assays to evaluate mutations in BRCA2342
• Genomic profiles of breast cancer342
• Role of molecular diagnostics in management of breast cancer343
• Tests for prognosis of breast cancer347
• Prediction of recurrence in breast cancer for personalizing therapy348
• Cervical cancer350
• Colorectal cancer350
• Diagnosis of hereditary nonpolyposis colorectal cancer350
• Detection of familial adenomatous polyposis coli351
• Diagnosis of colorectal cancer from DNA in stools351
• Detection of circulating tumor cells in colorectal cancer352
• Minimally invasive screening for colorectal cancer352
• Guanylyl cyclase C tests for colorectal cancer352
• Early diagnosis of colorectal cancer from blood samples353
• Gastric cancer353
• Head and neck cancer353
• Hematological malignancies354
• Chromosome translocations354
• Flow cytometry in diagnosis of leukemia354
• Gene chip technology354
• Laboratory assessment of leukemia355
• Molecular probes356
• Minimal residual disease356
• Screening of gene mutations in chronic myeloproliferative diseases356
• Lung cancer357
• Melanoma359
• Ovarian cancer359
• Mutation of genes360
• Relevance of genetic testing to management of ovarian cancer360
• Serum biomarkers for early detection of ovarian cancer360
• Biomarkers of ovarian cancer360
• Concluding remarks on testing for ovarian cancer361
• Pancreatic cancer361
• Prostate cancer362
• Gene expression analysis of prostate cancer by microarrays362
• Huntingtin Interacting Protein 1363
• Integrative genomic and proteomic profiling of prostate cancer363
• LCM for diagnosis of prostate cancer363
• PCA3 gene detection in urine364
• PCR assay for assessing silencing of protein cadherin 13 gene364
• Prostate biopsy for detection of prostatic intraepithelial neoplasia365
• Screening of multiple SNPs for risk of prostate cancer365
• Semen testing for prostate cancer biomarkers365
• Serum-protein fingerprinting in prostate cancer366
• Thyroid cancer366
• Gene expression biomarkers of thyroid cancer367
• Multiple endocrine neoplasia type 2B as risk factor for thyroid cancer367
• miRNA expression profiling in thyroid cancer367
• Urinary bladder cancer368
• Role of molecular diagnostics in the management of cancer368
• Risk assessment and prevention of cancer368
• Role of molecular diagnosis in the design of future cancer therapies369
• Molecular classification of cancer369
• Determination of cancer prognosis370
• Prognosis by tumor classification370
• Prognosis by cancer gene expression370
• Selection of anticancer drugs based on molecular diagnosis370
• Integrated genome-wide analysis of cancer for diagnosis and therapy371
• Personalized therapy for cancer patients371
• Pharmacogenetics and cancer therapy371
• Molecular diagnostics as an aid to selection of cancer therapy372
• Drug resistance in cancer373
• Role of organizatons in molecular diagnosis of cancer373
• Role of NCI in molecular diagnosis of cancer373
• Molecular profiling of cancer373
• Cancer Genome Atlas374
• Cancer Genetic Markers of Susceptibility Project374
• Support for future research in molecular diagnosis of cancer375
• Role of the International Cancer Genome Consortium375
• Future prospects of molecular diagnosis of cancer376
• Companies involved in molecular diagnosis of cancer376

8. Molecular Diagnostics in Biopharmaceutical Industry & Healthcare 382

• Introduction382
• Molecular diagnostics in biopharmaceutical industry382
• Molecular diagnostic technologies and drug discovery382
• Molecular diagnostics and pharmacogenetics383
• Molecular toxicology384
• Gene expression studies385
• Toxicogenomics385
• Toxicoproteomics386
• Mitochondrial assays387
• MetaChip387
• Molecular diagnostics and pharmacogenomics388
• Applications molecular diagnostics in gene therapy389
• Use of PCR to study biodistribution of gene therapy vectors389
• PCR for verification of the transcription of DNA390
• In situ PCR for direct quantification of gene transfer into cells390
• Detection of retroviruses by reverse transcriptase (RT)-PCR390
• Assessment of safety issues of gene transfer390
• Quantitative PCR for monitoring the effectiveness of gene therapy390
• Use of FISH for analysis of adeno-associated viral vector integration390
• Monitoring of gene expression by green fluorescent protein391
• Detection of microbial contamination in biopharmaceutical manufacturing391
• Role of PCR in detecting contamination of biopharmaceuticals391
• Contamination of biopharmaceuticals with prions392
• DNA tagging for control and tracing of drug distribution channels392
• Molecular diagnostics for organ transplantation392
• Tissue typing392
• Commercial products for transplant molecular diagnostics394
• Post-cardiac transplant patient monitoring for rejection396
• Blood Transfusion Screening396
• Molecular tests for screening of blood supply for viruses397
• Commercial molecular diagnostic technologies for blood screening398
• Bridge Amplification Technology399
• COBAS AmpliScreen HCV and HIV Assays399
• INACTINE399
• NucliSens Extractor system399
• Pall' s enhanced Bacteria Detection System400
• PCR combined with algorithm method400
• Prions detection in human blood401
• PRISMR automated system401
• Procleix HIV-1/HCV Assay401
• West Nile virus detection in human blood402
• Limitations of molecular diagnostics for blood screening402
• Molecular epidemiology403
• Molecular epidemiology of genetic diseases403
• Monogenic versus polygenic disorders404
• Critical issues facing genetic epidemiology404
• Molecular epidemiology of infectious diseases404
• Methods and purposes404
• Emerging infections405
• Human vs. non-human infections405
• Genetics and susceptibility to infectious disease406
• Molecular epidemiology of cancer406
• Molecular epidemiology of p53 gene mutations407
• Molecular epidemiology of link between virus and cancer407
• Molecular epidemiology and cancer prevention407
• SNPs and molecular epidemiology407
• Molecular diagnostics for identification of food-borne pathogens407
• Introduction407
• Molecular diagnostic methods used in food-borne infections408
• Limitations of use of molecular probes in food analysis409
• Optical biosensor for detection of Listeria-contaminated foods409
• Companies with technologies for food pathogen detection410
• Transmissible spongiform encephalopathies (TSEs)411
• Molecular diagnosis of TSEs411
• Companies involved in developing molecular diagnostics for TSEs414
• Detection of genetically modified organisms in food415
• Molecular diagnostics for detection of doping in sports415
• Screening of synthetic glucocorticosteroids in human urine416
• Detection of gene doping416
• Role of molecular diagnostics in future healthcare416
• Translation of genomic research into genetic testing for healthcare417
• Molecular diagnostics and disease management417
• Role of genetic biomarkers in disease management418
• Role of molecular diagnostics in personalized medicine418
• Integrated healthcare418
• Screening418
• Early diagnosis419
• Prevention419
• Therapy based on molecular diagnosis419
• Monitoring of therapy419
• Advantages and limitations of integrated healthcare420
• Commercially available systems for integrated healthcare420
• Combination of diagnostics and therapeutics421
• Companion diagnostics421
• Companies involved in companion diagnostics421
• Point-of-care diagnosis423
• Technologies for point-of-care diagnosis424
• Biochips for point-of-care diagnosis425
• Companies developing point-of-care diagnosis426
• Advantages versus disadvantages of point-of-care diagnosis428
• The impact of molecular diagnostics on clinical laboratory practice428

9. Molecular Diagnostics in Forensic Medicine and Biological Warfare 430

• Application of molecular diagnostics in forensic medicine430
• Technologies430
• Extraction of DNA from forensic samples430
• Mitochondrial DNA (mtDNA) analysis431
• Polymorphic Alu insertions431
• Single Nucleotide Polymorphisms (SNP) analysis432
• Short tandem repeat (STR)432
• Fluorescent detection systems433
• ABO genotyping433
• DNA analysis for identification of ancient or historical specimens433
• Applications433
• Applications in criminology434
• Identification of remains of military personnel435
• Identification of remains of victims of mass disasters435
• Parentage testing436
• Gender determination437
• Companies developing molecular diagnostics for forensic science438
• Molecular detection of biological warfare agents439
• Introduction to biological warfare agents439
• Role of PCR in the diagnosis of biological warfare agents440
• Multiplex PCR microarray assay to detect bioterror pathogens in blood440
• Laboratory diagnosis of Anthrax441
• Challenges in diagnosis of biological warfare agents441
• US government efforts for detection of biological warfare agents442
• The US Army Medical Research Institute of Infectious Diseases442
• Homeland Security Advance Research Projects Agency443
• Commercial development of diagnostic devices for biological agents443
• Companies developing diagnostic devices for biological agents443
• Biodefence microarray446
• Identification of genetic markers of individual pathogens446
• Microbial Identification System based on OptiChip"!447
• Hand-Held Advanced Nucleic Acid Analyzer447
• Nanogen' s portable detection device447
• Nanode Array Sensor Microchips448
• MicroChemLab448
• BioThreat Alert Test Strip448
• Benchtop living cell biosensor449
• BioForce NanoArray sensor technology449
• QTL handheld biosensor449
• Analyte 2000 biosensor450
• Airborne bacterial spore detection technology450
• Destruction and detection of anthrax by lysin450
• Biosensor based on mass spectrometry of microorganisms' s RNA450
• Bead ARray Counter451
• ProteinChip-based detection of bioterroism agents451
• TIGER biosensor451
• The PathAlert™ Detection System452
• VereThreat"!452
• Concluding remarks about biodefense applications of diagnostics452

10. References 454

Tables

• Table 1 1: Landmarks in development of molecular technology and its application to diagnosis23
• Table 1 2: Applications of molecular diagnostics42
• Table 2 1: Companies with products for nucleic acid isolation51
• Table 2 2: Some commercially available real-time PCR systems54
• Table 2 3: Applications of real-time PCR55
• Table 2 4: A selection of companies with commercially available FISH diagnostics80
• Table 2 5: Selected companies with RNA diagnostic tests81
• Table 2 6: Companies involved in whole genome amplification89
• Table 2 7: Companies involved in sequencing92
• Table 2 8: Comparison of methods of identification of unknown DNA sequences94
• Table 2 9: Classification of methods of gene expression analysis95
• Table 2 10: A selection of companies with gene expression technologies100
• Table 2 11: Companies involved in developing PNA diagnostics103
• Table 3 1: Applications of biochip technology in relation to molecular diagnostics109
• Table 3 2: Companies developing whole genome chips/microarrays115
• Table 3 3: Companies involved in biochips for molecular diagnostics116
• Table 3 4: Companies developing microfluidic technologies120
• Table 3 5: Biosensor technologies with potential applications in molecular diagnostics121
• Table 3 6: Important applications of biosensors131
• Table 3 7: Companies involved in application of biosensors in molecular diagnostics131
• Table 3 8: Selected labels for nucleic acid detection133
• Table 3 9: Selected companies with fluorescence and chemiluminescence products134
• Table 3 10: Companies involved in molecular beacon manufacture and research136
• Table 3 11: Selected companies involved in molecular imaging143
• Table 3 12: Nanotechnologies with potential applications in molecular diagnostics144
• Table 3 13: Companies developing nanomolecular diagnostics152
• Table 4 1: Applications of protein biochips/microarrays163
• Table 4 2: Companies involved in developing diagnostic applications of protein biochips170
• Table 4 3: Disease-specific proteins in the cerebrospinal fluid of patients172
• Table 5 1: Mutation detection technologies181
• Table 5 2: Technologies for SNP analysis188
• Table 5 3: A sampling of companies involved in technologies for SNP genotyping198
• Table 5 4: Application of preimplantation genetic diagnosis in monogenic disorders207
• Table 5 5: Companies involved in prenatal/preimplantation diagnostics209
• Table 5 6: CFTR genotyping in cystic fibrosis - companies and technologies210
• Table 5 7: X-linked immunodeficiency disorders214
• Table 5 8: Genes that cause cardiovascular diseases220
• Table 5 9: Available molecular diagnostics for neurogenetic diseases228
• Table 5 10: Companies offering genetic screening tests directly to consumers235
• Table 6 1: Molecular techniques for the diagnosis of infections237
• Table 6 2: Bacteria and fungi that can be detected by recombinant DNA tests250
• Table 6 3: Viruses that can be detected by recombinant DNA methods261
• Table 6 4: Companies with molecular diagnostics for avian influenza virus H5N1283
• Table 6 5: Companies developing POC tests for the diagnosis of infections298
• Table 6 6: Selected companies involved in molecular diagnosis of infections301
• Table 7 1: Estimated new cases of cancer in the US at most involved organs - 2008303
• Table 7 2: Tumor suppressor genes, their chromosomal location, function, and associated tumors304
• Table 7 3: Viruses linked to human cancer306
• Table 7 4: A classification of molecular diagnostic methods in cancer308
• Table 7 5: Desirable characteristics of biomarkers for cancer311
• Table 7 6: Approved monoclonal antibodies for cancer diagnosis320
• Table 7 7: Methods for comparison of gene-expression profilling in tumor specimens322
• Table 7 8: Impact of in vivo molecular imaging of cancer on oncology practice334
• Table 7 9: Companies developing cancer molecular diagnostics375
• Table 8 1: Applications of molecular diagnostics in the biopharmaceutical industry381
• Table 8 2: Molecular diagnostic technologies for drug discovery382
• Table 8 3: Molecular diagnostic technologies used for pharmacogenetic studies382
• Table 8 4: Companies with novel molecular toxicology technologies383
• Table 8 5: Applications of molecular diagnostics in gene therapy388
• Table 8 6: Companies involved in transplant molecular diagnostics393
• Table 8 7: Companies involved in molecular diagnostics of blood transfusions397
• Table 8 8: Pathogenic bacteria in food and targets for molecular diagnostic probes407
• Table 8 9: Companies involved in molecular diagnostics for food-borne infections409
• Table 8 10: Testing for harmful prions in brain tissue from dead cattle411
• Table 8 11: Companies involved in developing molecular diagnostics for TSEs413
• Table 8 12: Companies involved in companion diagnostics421
• Table 8 13: Applications of point-of-care diagnosis422
• Table 8 14: Companies developing point-of-care diagnostic tests425
• Table 9 1: Forensic and legal applications of molecular diagnostics432
• Table 9 2: Molecular technologies used for forensic applications437
• Table 9 3: Classification of biological and chemical agents used as weapons of mass destruction438
• Table 9 4: Biological warfare agents that can be identified by PCR methods439
• Table 9 5: Companies developing detection devices for biological warfare agents443

Figures

• Figure 1 1: Relation of molecular diagnostics to other technologies43
• Figure 2 1: Rolling circle amplification technology63
• Figure 2 2: A schematic view of the Invader operating system71
• Figure 2 3: Principle of fluorescent in situ hybridization75
• Figure 2 4: Repli-G system of Qiagen90
• Figure 2 5: DNA sequencing process91
• Figure 2 6: Electrochemical detection of DNA105
• Figure 2 7: Elements of a Scorpions primer106
• Figure 3 1: Affymetrix GeneChip technology110
• Figure 3 2: Basic principle of a biosensor121
• Figure 3 3: Surface plasmon resonance (SPR) technology126
• Figure 6 1: Use of DNA chips in diagnosing microbial infections239
• Figure 6 2: High throughput DNA pyrosequencing for pathogen discovery242

Part II

11. Ethics, Patents and Regulatory issues 11

• Introduction11
• Ethical concerns about genetic diagnosis11
• Ethical guidelines for molecular diagnostics12
• Ethical and regulatory aspects of direct-to-consumer genetic services13
• US public attitudes about genetic testing15
• Genetic testing for susceptibility to adult-onset cancer15
• Ethics of preimplantation genetic diagnosis16
• Preimplantation genetic diagnosis to screen for hereditary diseases16
• PGD to test for susceptibiliy to cancer16
• PGD and stem cells17
• Genetic research on stored tissues17
• Informed consent in clinical trials of in vitro devices17
• Concluding remarks about ethical issues18
• Insurance underwriting and gene tests18
• Should genetic information be available to health insurers?18
• A need for the re-examination of current views19
• Genetic Information Nondiscrimination Act of US19
• Patents for molecular diagnostics20
• PCR patents20
• Patenting DNA sequences20
• US policy on gene patenting relevant to molecular diagnostics20
• The impact of disease gene patents on molecular diagnostics21
• Licensing problems associated with genetic testing21
• Role of the WHO in genetic testing standards22
• Regulatory issues in the US22
• Assay Migration Studies for In Vitro Diagnostic Devices22
• Assessment of diagnostic accuracy23
• Sensitivity and specificity23
• Documentation of diagnostic accuracy23
• Assessment of laboratory-developed tests used by Medicare recipients24
• Discovery of incidental findings on genetic screening25
• Evaluation of companion diagnostics/therapeutic for cancer25
• FDA regulation of multivariate index assays26
• FDA guidance for IVDs to detect pathogens27
• FDA guidelines for devices to detect and differentiate HPV27
• FDA' s Microarray Quality Control28
• FDA and point-of-care diagnosis28
• Genetic testing of rare disorders29
• Quality control of molecular diagnostic laboratory procedures29
• Quality control of point-of-care tests30
• Regulation of IVD by the FDA30
• Regulation of in vivo diagnostics by the FDA31
• Regulation of analytic-specific reagents32
• Regulatory aspects of FISH32
• Regulation of genetic testing32
• Role of the FDA in genetic testing32
• Regulatory issues concerning blood and plasma products33
• United States Diagnostics Standards33
• Regulation of in vitro diagnostics in the EU34
• EU regulations for testing of blood products34
• Regulation of genetic testing in EU34
• Evaluation of diagnostic laboratory tests in the UK35
• Pre-implantation genetic diagnosis in the UK35

12. Markets for Molecular Diagnostics 37

• Introduction37
• Methods for study of molecular diagnostic markets37
• The overall market for diagnostic technologies38
• Molecular diagnostic markets according to technologies38
• Marketing strategies according to technologies39
• Nucleic acid isolation market39
• Market for PCR-based tests39
• DNA sequencing market39
• Cytogenetic market40
• Market for FISH technologies40
• Biochip/microarray market40
• Biosensor market41
• Nanobiotechnology for molecular diagnostics41
• Markets for gene expression technologies41
• Reagents and other disposable laboratory materials41
• Market for immunochemistry diagnostic42
• Markets for tissue diagnostics42
• Molecular diagnostic markets according to therapeutic areas42
• Genetic disorders43
• Prenatal testing44
• Cancer44
• Potential markets for cancer diagnosis according to type of cancer45
• Infectious diseases45
• Sexually transmitted diseases46
• Hospital-acquired infections47
• Testing for HIV drug resistance48
• Potential markets for avian influenza diagnostics48
• Cardiovascular diseases48
• Neurological disorders49
• Food testing49
• Screening of blood for transfusion49
• Tissue typing for transplantation49
• Marketing opportunities according to geographic areas49
• Unmet needs in molecular diagnostics50
• Major market trends51
• Markets according to home-brew and FDA-approved tests51
• Decentralization of molecular diagnostics52
• Point-of-care testing52
• Development of personalized medicine53
• Cost of sequencing the human genome53
• Cost of genotyping53
• Marketing companion diagnostics for personalized medicine54
• Development of low-cost tests54
• Simplification of test procedures55
• Increasing role of proteomics in clinical diagnostics55
• Forensic and legal applications55
• Marketing strategies55
• Role of alliances in commercialization of molecular diagnostics56
• Acquisitions vs collaborations56
• Analysis of collaborations in molecular diagnostics59
• Licensing of the technologies60
• Strategies related to laboratory facilities and technologies60
• Strategies relevant to the healthcare system61
• Cost-Benefit studies61
• Genetic susceptibility testing61
• Preventive medicine strategies62
• Targeting treatable and common diseases62
• Information/education62
• Physician education62
• Patient education63
• European diagnostic information platform63
• Regulatory strategies64
• Merger of in vitro and in vivo diagnostics64
• Integration of diagnostics with therapeutics64
• Diagnostic applications in clinical trials64
• Prospects for development of new technologies65
• Drivers for the development of molecular diagnostics65
• Factors slowing the development of molecular diagnostics65
• Government support of research relevant to molecular diagnostics66
• Cost of sequencing the human genome66
• European projects for improving molecular diagnostics68
• European Consortium for developing new DNA analysis tools68
• EU project for improvement of IVD tools procedures69
• Genetic knowledge parks in the UK69
• Molecular diagnostic opportunities in defense against bioterrorism69
• Molecular diagnostics for food safety70
• POC diagnostics for the developing countries70

13. Companies involved in molecular diagnostics 71

• Introduction71
• Major players in molecular diagnostics71
• Profiles of selected companies72
• Collaborations418

Tables

• Table 12 1: Share of in vitro diagnostics in the global diagnostic market 2008-201838
• Table 12 2: Molecular diagnostics markets according to technologies from 2008-201838
• Table 12 3: Molecular diagnostics markets according to applications 2008-201842
• Table 12 4: Markets in 2008 for tests to screen healthy persons for genetic disorders43
• Table 12 5: Markets in 2008 for molecular diagnostic tests for cancer44
• Table 12 6: Molecular diagnostic markets for selected cancers 2008-201845
• Table 12 7: Markets value in 2008 for molecular diagnostic screening for infections46
• Table 12 8: Future markets for HAI diagnostics 2008-201347
• Table 12 9: Molecular diagnostic markets according to geographical areas 2008-201850
• Table 12 10: Molecular diagnostic markets according to home-brew and approved tests51
• Table 12 11: Marketing strategies for molecular diagnostics55
• Table 12 12: Takeovers of molecular diagnostic companies56
• Table 12 13: Advantages of the integration of diagnostics with therapeutics63
• Table 13 1: Top ten players in molecular diagnostics71
• Table 13 2: Collaborations of companies in molecular diagnostics417

Figures

• Figure 12 1: Unmet needs in applications of molecular diagnostics51
• Figure 12 2: Proportion of various areas in molecular diagnostic collaborations59