QUALITY CONTROL & QUALITY ASSURANCE in Molecular Laboratory

Presented By Toheed Kausar

Introduction
Molecular Pathology
In AKUH , Molecular Pathology section was established in 1995 in Clinical Laboratory , performs DNA and RNA based testing to provide diagnostic information to physicians and clinicians .

Introduction
Molecular Pathology

Molecular Molecular Molecular Virology Oncology Genetics HCV PCR

QHCV HCV Genotyping HBV PCR QHBV HBV Genotyping HDV PVR QHDV HEV PCR CMV PCR HIV PCR QHIV Dengue IgM CCHF PCR HSV1 &2 PCR H1N1 PCR BCR PCR QBCR APL PCR JAK2 mutation FLT3 Mutation NPM1 Mutation

HLA Testing

Thalassemia PCR (CVS) T. HLA Typing Thalassemia PCR HLA DR (Blood) HLA Typ minus SMA PCR DR DMD HLA B27 PCR Delta 508 Mutation MTHFR Polymorphism PCR Q-Pre Q-post HbE Mutation HbD Mutation HbS Mutation DNA banking IL28B by PCR

Molecular Molecular Bacteriolog Parasitology yAFB PCR Naegleria

CTA Sp by PCR

Quality Assurance
Quality assurance is the series of checks and

balances that ensure laboratory results are accurate and reliable. The common components of the laboratory quality assurance program include pre analytical (specimen collection, handling, and storage), analytical (instruments/equipment, reagents, personnel) post-analytical (result reporting and interpretation) procedures. All quality assurance procedures must be written and included in the laboratory procedure manual with documentation of the performance of the

Quality Control

Quality control is an integral component of quality assurance and is the aggregate of processes and techniques to detect, reduce, and correct deficiencies in an analytical process.

QC & QA

Quality control is concerned with the end product, while quality assurance is processoriented.

Quality Management System (QMS) in Molecular Pathology

Quality Assurance QA

Quality Control QC

- Accuracy: - Precision: - Analytical specificity:

- Internal QC - External QC

Quality Control in Molecular Pathology

Controls are samples of known type or amount

that are treated like and run with patient specimens. Interpretation of test results always includes inspection of controls and standards to verify acceptable test performance. With qualitative tests, a positive, negative, internal and, in some cases, a sensitivity control are required. The sensitivity control defines the lower limit of detection for more meaningful interpretation of negative results.

QMS in Molecular Pathology

ObObjective (Sample Handling/Processing): jecample Receiving): 1.To ensure specimen meets required quality specifications. 2. To maintain identification and traceability of patient supplied products from time of its receipts till its test results. 3. To ensure traceability of patients/physicians supplied product at all processing and storage level from receipt, till dispatch of results.

QMS in Molecular Pathology

Objective(Sample Handling/Processing):

To ensure that necessary check / tests are carried

out and documented to verify that specific requirements for the document are met. To document and maintain the records for all inspection and testing. To dispatch specimen to appropriate laboratory sections for further processing. To ensure that approved procedure manual are available for consultation.

QMS in Molecular Pathology

Objective(Storage/Retention): To ensure that all products from point of receipt until dispatch of final product are controlled, properly documented, handled safely, stored as per Specific requirements to prevent from damage or deterioration of products.

QMS in Molecular Pathology

Instrument Maintenance: Instruments used in the molecular laboratory must be monitored and maintained for consistent performance and accurate test results. Reagents: When reagents are replaced in a test method, the new lot is ideally tested on a previously positive and negative specimen as well as the run controls.

QMS in Molecular Pathology

Reagents:
Instructions on the preparation of reagents and the

quantities used in each assay are included in the written laboratory protocol for each procedure. Lot numbers and working stocks of probes and primers used in amplification methods are documented and matched to test performance in the runs in which they were used.

QMS in Molecular Pathology

Chemical Safety: Volatile and flammable

reagents are stored in properly vented and explosion-proof cabinets or refrigeration units. Secondary or reinforced containers are required for transport handling of dangerous chemicals.

QMS in Molecular Pathology

OBJECTIVE (Result dispatch)

1. To ensure quality results.

2. To make certain that reports are released as

per contract. 3. To ensure documentation of verbal communication of urgent results.

Important Definitions
Analytic sensitivity: lower limit of detection of the analyte(The t(14;18) translocation test can detect 1 translocated cell in 10,000 normal cells, an analytic sensitivity of 0.01%.) Clinical sensitivity: Ability of test result to predict a clinical condition (95 of 100 patients with a gene mutation have a disease state, a clinical sensitivity of 95%.)

Important Definitions
Analytic specificity: Ability to detect only the analyte and not nonspecific targets. (The Invader assay for factor V Leiden successfully detected mutations in 18 positive specimens while yielding negative results for30 normal specimens (no false positives.) Clinical specificity: Disease-associated results only in patients who actually have the disease conditions. (1 of 100 normal specimens displayed a gene mutation (1 false positive) a clinical specificity of 99%.)

Important Definations
Precision: Reproducibility of test results.

A quantitative method yields 99 results in agreement out of 100 runs, a precision of 99%. Analyte measurement: The range within which a specimen may be measured directly (without dilution or concentration). A qPCR HSV assay yields reproducible linear results from 10 to 107 copies of HSV per 20 ul of CSF. Specimens within this range are measured directly.

Important Definitions
Analytic accuracy: Production of correct results.(

99 of 100 specimens with mutations in the BG gene are detected by sequencing with no mutations detected in normal specimens.)

College of American Pathologist (CAP)

College of American Pathologists is a medical

society serving more than 17,000 physician members and more than 6,000 laboratories are accredited by CAP throughout the world. It is the worlds largest association composed exclusively of pathologists and is widely considered the leader in laboratory quality assurance. It is the best Laboratory Accreditation Program which is useful for demonstrating and improving quality. It certifies that the lab is committed to quality patient care and achieving excellence.

JOINT COMISSION INTERNATIONAL ACREDITATIN (JCIA)

The Joint Commission, formerly the Joint

Commission on Accreditation of Healthcare Organizations (JCAHO), is a private sector United States-based not-for-profit organization. The Joint Commission operates accreditation programs for a fee to subscriber hospitals and other health care organizations.

JOINT COMISSION INTERNATIONAL ACREDITATIN (JCIA)

The company updates its accreditation standards

and expands patient safety goals on a yearly basis, and posts them on its Web site for all interested persons to review, making this information and process transparent to all stakeholders ranging from institutions, to practitioners, to patients and their advocates.

INTERNATIONAL ORGANIZATION FOR STANDARDIZATION (ISO)

The International Organization for Standardization

(ISO) is a non-governmental organization based in Geneva, Switzerland, that works to develop technical standards for products and services all around the world.

Summary
Proper specimen handling is required for

accurate test results. Specimens should be held and stored under conditions that will preserve nucleic acids. Molecular test performance is monitored by the use of quality controls. Instruments should be maintained and calibrated for accurate detection and measurement of analytes. Reagents are prepared, stored, and used as recommended by manufacturers and/or laboratory protocol. Raw data should be documented and results clearly reported.