molecular laboratory must have an ongoing bio-safety sop and also quality improvement program to...
TRANSCRIPT
Molecular Laboratory must have an ongoing Bio-safety SOP and also quality improvement program to
monitor and evaluate objectively and systematically the quality and
appropriateness of the information/service it provides
Why do we need safety
Safe handling of hazard microorganisms, Chemicals and radiations
Safe conditions for the work to keep the tests in right results
Bio-Safety for the Personnel
Hazard Microorganisms (Risk Group 1 to 4) Chemicals Radiations
General Bio-Safety Rules Specific Considerations
Ethidium Bromide, Acrylamide, UV radiation (Door Glasses)
Specimens
Specimens are to be handled according
to SOPs, with special attention to any safety precautions (Laboratory General Bio-Safety)
Hazard Microorganisms
Each detectable organisms must be classified on its Bio-Safety Level
Risk of Lab. Infections must be determined for each organisms according to applied diagnostic tests
Blood-borne and respiratory pathogens
Hazard MicroorganismsContinue
If a dead air UV hood is used, turn on the UV lamp at least 15 minutes prior to working
When using a biological safety cabinet (BSC), it must be on 1/2 hour prior to specimen preparation procedures.
Working surfaces must be decontaminated prior to specimen preparation with 10% bleach solution (freshly made daily) followed by 70% ethanol rinse
Gloves must be worn at all times for your safety when handling specimens
Specimen preparation area
Defined Flow work and Lab. Design
Awareness to the Applied Lab. Procedures
(provide necessary SOP)
Specimens In general, specimens should be received and accessioned in an
area of the facility, that is isolated from testing areas.
separation and aliquoting of original specimens
Back up specimens should be kept frozen at -20oC to - 70oC.
and avoided of freeze-thawing Use of sterile container
Patient Specimens and Genomic Materials should be stored separately than
reagents
Receiving samples:
Safe conditions for the work Continue
Gloves are to be changed frequently, anytime you contaminate them, whenever a specific procedure SOP dictates, and never be worn outside Post Amplification Area, especially in Pre Amplification Area
Aerosol barriers tips are used for pipetting into amplification tubes and pipetting amplification product
Other routes of contamination
Contamination of negative specimens with control specimens
False negative results due to inhibitors transferring from environment
(Such as Latex gloves….) and inefficient extraction methods
(Accreditation Protocols)
Molecular Lab.
Specific flow work and Lab designDepends on applied Technique
Separation of work areas: Pre-Amplification area, Post-Amplification area, Storing space Specific instruments, materials and other
condition for each area
How contamination can be distributed
Routs of contamination depend on the technique
target amplification
signal amplification
Decontamination
Post-amplification products from previous analysis can be distributed
Decontamination and inactivation protocols for the test and environment
Work-benches and sinks should be decontaminated each day of use
Products must be discard in a safe way
General Consideration in Designing Molecular Lab.
Each sample test contains 1010-1012 amplicons, distributed easily by hand, hair, Lab coat, samplers,…….
Reagents, materials and instrument can be contaminated easily
Complete separation between pre & post amplification areas must be considered
Continue
Air flow should be from clean area to dirty area Special program must be followed if different
PCR sets are being used Additional materials recommended to divide in
small sections and stored in separate space from PCR Lab
General Rules
Each Working space must have positive pressure in Pre-Amplification Area
Transferring reagents, materials, and any instrument must be avoided from dirty area to clean area
Lab coat and gloves must be replaced before entering to the clean area
Necessary material must be kept at each lab Each lab must have UV, Bleach (10%)
Preparation Lab
Preparation Lab is the center of the PCR Lab Each solution must be checked carefully Lab stocks must be divided to small amount
and used one by one in the lab Just mixture reaction will be prepared in this lab
without any DNA samples Preparation Lab must have its own specific
materials, samplers, buffers, and tube samples
Extraction room Any extraction must be carried out under safety
cabinet Necessary buffers, and materials must be taken out
from freeze and kept at the room temperature before work
It is recommended of two gloves with working of hazardous specimens
10% bleach are recommended to used for any probable contamination of centrifuge, safety cabinet, and so on
Sampler tips must be un-reusable and plug tips recommended
Amplification room
Thermocycler must be kept in a small clean area
Different control negative must be considered in order to insure the quality of the test (buffers, primers, dNTP, …)