Molecular Pathology - Part 1
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| Revolution in which area of science is having a profound impact on the field of anatomic and clinical pathology (diagnostic medicine). | Molecular biology
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| Techniques capable of detecting DNA in very small quantities in tissue and body fluids, enabling detection of various disease states, including cancer. | Polymerase chain reaction and other amplification and hybridization methods
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| Utility of Robust high-throughput and highly sensitive genetic methods created by human genome project | Detecting abnormal genes in patient specimens and for establishing patterns of gene expression that are characteristic of specific diseases.
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| Utility of Proteomic methods | Detecting protein expression in serum samples from patients for diagnosis of different types of cancers.
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| Methodologies that require sophisticated mathematical methods of pattern recognition that allow for detection of disease. | Gene array high throughput sequencing and proteomic methodologies
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| Skills required to understand molecular diagnostic testing | Familiarity with the basics of nucleic acid biochemistry and biology
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| Differences in stability of DNA and RNA | The chemical stability of double-stranded DNA stands in contrast to the lability of RNA.
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| Basis for DNA replication, RNA transcription, and hybridization assays | Base pairing of nucleic acids dictated by energetically favorable rules
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| Analysis impact of Chemical similarity of nucleic acid molecules, regardless of their source | Methods of extraction, storage, and handling are similar
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| Enzymes that synthesize and modify nucleic acids | Harnessed as tools for molecular biology and molecular diagnostics (e.g., polymerases, transcriptases, nucleases, ligases)
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| Nucleic acid analyses include | Electrophoresis, hybridization assays, amplfication techniques, sequencing, and polymorphism detection
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| Complete assays or diagnostic tests combine several of Nucleic Acid Analysis techniques | Such as amplification, electrophoresis, and hybridization.
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| Is Molecular diagnostics now a part of the mainstream of laboratory diagnostics. | Yes. Nucleic amplification technology has created new opportunities for the clinical laboratory to affect patient care.
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| Most mature and widely used nucleic acid amplification method | PCR
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| What is the milestone in biotechnology that heralded the beginning of molecular diagnostics. | Development of the PCR by Mullis and colleagues (Saiki et al, 1988)
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| Methods other than PCR with diagnostic applications are based on | These are based on target, probe, or signal amplification
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| Array Technology | Provides a unique and powerful approach to screen a sample for dozens to thousands of genes. Current array fabrications allow for diverse platforms with better detection and reduced cost for high-density applications.
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| When is a positive signal generated in Microarray | Positive signals are generated when a tagged nucleic acid moiety hybridizes with its complementary probe localized on a solid support (i.e., “chip”).
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| Proposed applications of array technology | Molecular staging of tumors to the identfication and characterization of microbial agents. (Care must be taken when performing and interpreting microarray experiments, because numerous factors must be considered in analysis.)
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| Factors making array technology more commonplace and user friendly. | Improvement of quality and yield of nucleic acid extraction from fixed and frozen tissues combined with continued miniaturization and cost reduction of platforms
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| How does Identification of chromosomal abnormalities aid in clinical diagnosis and treatment. | This often correlates to disease and phenotypic abnormalities and thus will
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| What information can Identification of a cytogenetically abnormal clone in a cancer can provide | Information on diagnosis, prognosis, treatment, and disease progression.
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| Technologies based on establishment of Standard morphology and a specific banding pattern have been established for all human chromosomes | Karyotype analysis, fluorescence in situ hybridization (FISH), microarray analysis, and DNA sequencing, are used to uniquely identify chromosomes and determine if anomalies exist.
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| The two basic categories of cytogenetic abnormality are | Numerical and Structural Cytogenetic abnormalities
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| Examples of syndromes that have been specifically linked to particular chromosomal anomalies | Trisomy 21 in Down syndrome; 45,X in Turner syndrome; and deletion of 22q11.2 in velocardiofacial syndrome.
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| Cytogenetic abnormalities types and detection age | TYPES - de novo or inherited.
DETECTION - may occur at any stage of a person’s life from prenatal to adulthood.
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| Fluorescence in situ hybridization (FISH) | A targeted assay that combines technologies from both cytogenetics and molecular genetics and can provide important diagnostic information about specific genes.
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| Cytogenetic analysis | Can be used in nearly every medical specialty and is an important element in clinical laboratory medicine.
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| Microarray analysis and DNA sequencing | Have identified new genetic syndromes. These technologies will be key to optimizing genetic testing in the future.
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| How to handle increased volume of molecular testing for infectious diseases | Utilizing Automation
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| Utility of Molecular testing in cancer | Diagnostics, prognostics, and therapeutic decision making.
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| Who all have impact of Molecular testing for inherited disorders | Both the patient and family members.
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| Laboratories performing testing on human specimens for the purposes of diagnosis, prevention, or treatment of diseases are subject which regulations | Federal regulation imposed by the Clinical Laboratory Improvement Act of 1988 (CLIA ’88) and amendments of 2003.
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| What is the Genetic Information Nondiscrimination Act of 2008 (GINA) | Federal law that prohibits discrimination in health coverage and employment based on genetic information.
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| What is Analyte-specific reagent (ASR) | It is a term devised by the FDA applicable to reagent(s) that may be components(s) of an LDT or IVD assay.
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| Lab Requirements of molecular testing methodologies involving amplification with high potential sensitivity | Strict control of air flow, movement of personnel, and physical separation of laboratory spaces devoted to pre- and post-amplification.
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| Requirements for Laboratory directors, technical supervisors, and molecular technologists of a CLIA-approved laboratory | Qualifications per CLIA and other regulations as applicable.
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| Changes in Process Reimbursement for molecular diagnostic tests | The process of determining coverage, coding, and payment is adapting to emerging technologies.
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| Current legal position of patenting naturally ocurring DNA | The Supreme Court has determined that naturally occurring DNA is not patent eligible.
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| Impact of Emerging genomic technologies with associated bioinformatics pipelines | Increasing demands on laboratory information systems.
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| The menu of molecular pathology laboratories may vary significantly and is determined by which three principal factors | Clinical requirements and usefulness, Laboratory competency, and Test costs.
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| What should Quality assurance programs monitor | All phases of testing to include preanalytical, analytical, and postanalytical phases. The details will depend on the assay but may include monitoring of turnaround time, proficiency testing, equipment performance, and personnel continuing education.
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| What is included in Assay design and development | Assessment of clinical validity, verification of assay performance, validation, and assessment of both analytical and clinical performance characteristics.
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| What testing phases should be included in Quality control programs | All phases of analytical testing to cover performance monitoring of reagents and instruments.
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