Chapter 11.

LABORATORY SAFETY

BIOSAFETY

Safety is the responsibility of everybody working in the laboratory and safe procedures
must be adhered to at all times. The following statements provide some basic rules for
safety in the laboratory.

! Blood and body fluid precautions are to be consistently used with all clinical
specimens of blood or other potentially infectious material (Universal Precautions).

! Use barrier protection at all times (laboratory coats, gloves, or other appropriate
barriers).

! Good laboratory practices should be followed. Eating, drinking, or smoking is not

permitted in the laboratory.

! Mechanical pipetting devices are to be used for all liquids in the laboratory. Mouth
pipetting is dangerous.

! Biosafety Level 2 practices should be followed when handling all specimens. Class I
or II biological safety cabinets or other physical containment devices should be used
for all manipulations of agents that cause splashes or aerosols of infectious materials.

! Adequate and conveniently located biohazard containers for disposal of contaminated

materials should be available.

! Countertops and surfaces of biological safety cabinets should be wiped with a

disinfectant (0.5 sodium hypochlorite is preferred) routinely after working with
infectious agents or clinical specimens.

! Wash hands often -- Especially before leaving the laboratory and before eating.
Protective clothing should be removed before leaving the laboratory.

HAZARDOUS CHEMICALS

Acetone will be used for fixation of cell cultures in the microneutralization assay.
Caution: acetone is extremely flammable as liquid and vapor, and harmful if inhated.
Eye and Skin contact: Immediately flush eyes with plenty of water; get medical
attention
Ingestion: Get medical attention
Inhalation: Remove to fresh air

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Hydrogen peroxide will be used as substrate for the HRPO-conjugate in
immunoperoxidase staining.
Caution: hydrogen peroxide may cause severe irritation of skin, eyes, and mucous
membranes and respiratory irritation.
Eye and Skin contact: Flush with plenty of water; get medical attention.
Ingestion: Give large volumes of water or milk if conscious;
DO NOT INDUCE VOMITING; get medical attention.
Inhalation: Remove to fresh air

Sodium azide is added as a preservative in a concentration of 0.1% to some of the

reagents included in the WHO Influenza Reagent Kit and the monoclonal antibodies.
Caution: sodium azide may be fatal if swallowed, inhaled, or absorbed through skin.
Eye and skin contact: Flush with plenty of water, remove contaminated clothing
Inhalation: Remove to fresh air, give artificial respiration or oxygen if required.
Ingestion: Give large quantities of water if conscious and not convulsive. Induce
vomiting; get medical attention.

Chloroform is used in the alternate methods for RNA extraction and the dsDNA
purification.
Caution: Chloroform is a possible human carcinogen and may be fatal if swallowed
or inhaled. Inhalation or ingestion may cause nausea, vomiting, headache, or
dizziness. Eye contact may cause corneal damage. Skin contact may cause irritation
or dermatitis.
Eye and skin contact: Flush with water for at least 15 minutes.
Ingestion: If swallowed and conscious, give large amounts of water. Induce
vomiting. Call a physician.
Inhalation: Remove to fresh air. Give oxygen if breathing becomes difficult. Call a
physician.

Ethidium Bromide is an intercalating dye that will be used to visualize DNA in agarose
gels.
Caution: Ethidium bromide is a powerful mutagen and is moderately toxic. Gloves
should be worn when working with solutions containing this dye, and a mask should
be used when weighing it out.
Eye and skin contact: Flush with water for at least 15 minutes.
Ingestion: Call physician immediately. Do not induce vomiting unless instructed by
medical personnel
Inhalation: Remove to fresh air. Give oxygen if breathing becomes difficult. Call a
physician.

Phenol is used in the alternate methods for RNA extraction and dsDNA purification.
Caution: Phenol is highly corrosive and can cause severe burns. This agent is rapidly
absorbed through the skin. Wear gloves, lab coat, and safety glasses when handling
phenol. All manipulations should be carried out in a chemical hood.
Eye and skin contact: Rinse with large volumes of water and wash with soap and
water. Do not use ethanol.

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Ingestion: If conscious, give activated charcoal in water, olive oil, or margarine; then
induce vomiting. Call a physician.
Inhalation: Remove to fresh air. Give oxygen if necessary. Call a physician.

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