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

Safe conditions for the work to keep the tests

in right results

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, …)

Detection Lab.

Test tubes can be opened after amplification just in Detection room

To prevent the distribution of amplified products 10% bleach and UV should be available

It is recommended not to keep the remaining products after any experiment

Lab coats and gloves must be removed