PRELIM LAB 1: QUALITY ASSURANCE AND CONTROL Flashcards
wide range of techniques and methods used to study biomolecules (DNA, RNA, proteins, nucleic acids and other protein molecules)
MOLECULAR ANALYSIS
Main purpose: for diagnostic or investigative purposes
- SPECIFIC APPLICATION OF MOLECULAR ANALYSIS TECHNIQUES
- To define a treatment strategy for individual patient
- Should be made appropriately and should be offered in a timely manner to achieve accurate and effective results
MOLECULAR TESTING
Workflow Molecular Test
activities that occur BEFORE the actual analysis of specimen
PREANALYTICAL PROCESS
Workflow Molecular Test
ACTUAL TESTING and analysis of the specimen in laboratory
Ex: extraction of DNA and RNA in cell
ANALYTICAL PROCESS
Workflow Molecular Test
- activities that occur AFTER the analytical testing is complete and results is generated
POSTANALYTICAL PROCESS
errors encountered in preanalytical process
PREANALYTICAL ERRORS
An aliquot removed from a specimen is NEVER RETURNED to the original tube or vessel
T OR F?
TRUE
Collection Tubes for Molecular Testing
chemistry, serum, viral antibody
RED
Collection Tubes for Molecular Testing
molecular biology studies
YELLOW
Collection Tubes for Molecular Testing
Immunology, virology
GREEN
Collection Tubes for Molecular Testing
Virology, molecular biology studies
LAVENDER
Collection Tubes for Molecular Testing
cytogenetics studies & molecular studies
BROWN
Special Collection Tubes
- WBC separation upon configuration and also used for HIV, HCV analysis
Tubes designed to stabilize RNA.
PLASMA PREPARATION TUBE (PPT)
- Designed by Center Disease Control and Prevention (CDC)
- A SET OF INFECTION PRECAUTION PRACTICES to protect the transmission of infectious diseases
- Hand hygiene, PPE, safe handling of sharps
- Transmission based precautions: prevents airborne/contact transmissible agents
- Contact precautions: for direct patient care where there is the potential for direct exposure to infectious agents on or from the patient
STANDARD PRECAUTION
Common Decontamination Agent
Heat
Ethylene oxide
Glutaraldehyde
10 % Hydrogen peroxide
10% Formalin
5.25% Hypochlorite
Formaldehyde
Detergents
Phenols
UV radiation
Ionizing radiation
Photo-oxidation
developed a series of warning label for universal use on all chemical container
Ex: chemical has the following NFPA hazard
NATIONAL FIRE PROTECTION ASSOCIATION (NFPA)
documents that provide information about the properties, hazards, handling, storage and emergency measures for substances/mixtures
MATERIAL SAFETY DATA SHEETS (MSDS)
___________require validation of the performance of clinical test methods & reagents in accurately detecting/ measuring analytes prior to use in human testing
FOOD AND DRUG ADMINISTRATION (FDA)
Outcome where the model correctly predicts the positive class
TRUE POSITIVE
Outcome where the model correctly predicts negative class
TRUE NEGATIVE
Outcome where the model incorrectly predicts the positive class
FALSE POSITIVE
Outcome where the model incorrectly predicts the negative class
FALSE NEGATIVE
- CHANGE IN ASSAY RESPONSE with corresponding change in analyte
Ex: All positive reference standard tested positive with the new assay and the analytical sensitivity of the assay is 100 % - TP/ TP + FN × 100
ANALYTICAL SPECIFICITY
ANALYTICAL ACCURACY
ANALYTICAL SENSITIVITY
ANALYTICAL SENSITIVITY
- ABILITY TO DETECT ONLY THE ANALYTE and not nonspecific targets
Ex: The invader assay for factor V Leiden successfully detected mutation in 18 positive specimen while yielding negative results (for 30 normal specimens
TP/TN + FN × 100
ANALYTICAL SPECIFICITY
ANALYTICAL ACCURACY
ANALYTICAL SENSITIVITY
ANALYTICAL SPECIFICITY
- PRODICTION OF CORRECT RESULTS
Ex: Of 100 specimens with mutations in HCM gene, 90 are detected by sequencing with no mutations detected in normal specimens - TN + TP/TN + TP + FN + FP × 100
ANALYTICAL SPECIFICITY
ANALYTICAL ACCURACY
ANALYTICAL SENSITIVITY
ANALYTICAL ACCURACY
- Refers to external specimen from reference source supplied to independent laboratories
- Determines the performance individual laboratory for specific tests/measurement and is used to monitor laboratory continuing performance
- Performed at least 2x a year
CONTROLS
PROFICIENCY TESTING
VALIDATION
PROFICIENCY TESTING
- Sample of known type/amount that are treated like and run with patient specimen
- Serve as reference points to validate and monitor various steps of molecular experiments, ensuring the reliability of and accuracy of results
CONTROLS
PROFICIENCY TESTING
VALIDATION
CONTROLS
- Range of activities that enable laboratory to achieve high level of accuracy and proficiency
- Periodic review and documentation of test result is required
- Monitored controls and conditions are reviewed monthly at determined intervals to document outliers/systems errors.
QUALITY ASSURANCE
- Systematic process and procedures to monitor, assess and ensure the accuracy, precision and reliability of molecule assays and experiments
- Use positive, negative and sensitivity control to monitor assay performance
QUALITY CONTROL MEASURE
- Essential document that provide detailed instruction on how to perform specific task/processes
- Should be clear, detailed, and readily accessible to all laboratory personnel
DOCUMENT SOP
- Through training for laboratory staff on SOPs, protocols, and safety procedures
- Regular assessment of competency personnel (ex: proficiency testing and training programs)
TRAINING AND COMPETENCY ASSESSMENT
Detect a specific target (cell surface protein/DNA mutation
Analytic Specific Reagents (ASRs)
OR
In vitro Diagnostic Devices/Reagents
Analytic Specific Reagents (ASRs)
for use in the diagnosis of disease, products that may be to collect, prepare and examine specimen
Analytic Specific Reagents (ASRs)
OR
In vitro Diagnostic Devices/Reagents
In vitro Diagnostic Devices/Reagents
Result verification
Data interpretation
Result reporting
Storage of sample and data
Archiving
POSTANALYTICAL PROCESS
DNA/RNA extraction
DNA/RNA Quantification
Purity Determination
ANALYTICAL PROCESS
Isolating nucleic acids (DNA/RNA from biological samples
DNA/RNA extraction
Characterization of nucleic acids
DNA/RNA Quantification
Assessing the presence of contaminants (proteins or organic compounds) in nucleic acid samples.
Purity Determination
Ability of test result to predict a clinical
condition
Clinical sensitivity
Least detectable presence of the analyte
Detection limit, limit
of detection
Disease-associated results only in patients
who actually have the disease conditions
Clinical specificity
Agreement between independent test
results
Precision
Consistency of test results produced from
the same procedure
Reproducibility
The range within which a specimen may be measured directly (without dilution or
concentration
Analyte measurement range
The range within which test results are considered to be valid (with or without
dilution)
Reportable range
Expected analyte frequency or levels from
a population of individuals
Reference range
Quantitative correlation between test result and actual amount of analyte
Linearity
CONTROLS:
defines the lower limit of detection for more meaning interpretation of negative results
sensitivity control
distinguishes true-negative amplification results from false negatives resulting from amplification failure.
amplification control
