Guidelines for making SOP to be followed in Polio Virus Isolation and Identification

Purpose: This SOP described how to avoid hazardous procedure like breakages, leaks and spills during the Work. Avoiding the use of defective should minimize breakages, Leaks and spills, Cracked,    Chipped containers by firm closures or sealing and by placing tubes and bottles in racks for support

A – BREAKAGE OF CELL CULTURE BOTTLES OR TUBES

v Cuts due to breakage, which handling cell cultures, particularly which stoppering tubes are common in cell culture, so when it should be happen tubes should be flawless or alternatively screw – cap may be used. If the tubes containing media or any other infectious materials are broken by accident or any how first take all that piece of glass tubes very carefully with gloves and discard it in to the 1% Sod. Hypochlorite solution.

B – PIPETTING

v Fluid transfers can cause dangerous aerosols and spills. Vigorous pipetting causing frothing & bubbling of fluids must be avoided.

v To avoid aerosols cause by pipetting fluid manipulating should be slow and gentle using rubber teat or bulb or automatic pipette

C – GRINDING AND HOMOGENIZATION

v Aerosols also cause by vigorous shaking and during chloroform treatment and also grinding by sample processing.

v Aerosols are generated during these procedures which should preferably by carried out using class1 BSC.

v After homogenization a period for the aerosol to settle should be allowed to elapse before opening the apparatus.

D - CENTRIFUGATION

v The potential hazard in centrifugation is obvious i. e. aerosol dispersion due to leakage of over filled or insecurely capped containers or their breakage due to the use of flawed containers or careless balancing or contents.

v Avoid these should be happens any time of breakages or the inside the centrifuge. Take out the tubes or pieces of tubes and discard it in to discarding pan containing freshly prepared 1% Sod.

v Also take out the adaptor of this breakage of tubes and dip into the container of freshly prepared 1% Sod. Hypochlorite solution.

SAFETY CONDITION

1)     To avoid any type, of accident in to the lab. Care should be taken during every working session.

2)     Every person must be wearing a gown and gloves for self-protection due-ring any type of work in to the lab.

3)     No exposed infected items reach the disinfections/ autoclaving / incineration departments.

4)     All open items should be securely closed or enclosed and sharp instruments sheathed or enclosed in lidded receptacles

5)     Refrigerated centrifuge and shaker is cleaned with 1% Hypochlorite solution and then cleaned with autoclaved D.D.W.

6)     Start up and shut down register of all BSC are maintained.

1)     All infectious waste are discarded in 1% Hypochlorite solution and then sterilized by physical method (Autoclave) and then sent to incinerator daily.

2)     Clean the hood with 1% Hypochlorite solution, close the shutter and turn on U.V. light and air flow for at least 10min. between each work.

5.1 Laboratory accidents-Chemical

Purpose: This SOP explains how to protect from chemical hazards.

Materials: All chemicals, which are used in the laboratory.                       

GENERALS DESCRIPTION:

v Storage of chemicals

·       Only minimum amounts of the chemicals should be stored in the laboratory for daily use.

·       Flammable substances must be kept away from any other chemicals. e.g. Spirit is stored in cupboard in museum room. Chloroform is stored in room no.20 which is locked.

·       All bottles containing chemicals like strong acids and alkalis should be at floor level in drip trays.

v Incompatible chemicals                           

Many common laboratory chemicals react in a dangerous manner if they come into contact with one another. The following chemicals are kept at a distance from each other.

·       Acetic acid: With Chromic acid, Nitric acid, Hydroxyl compounds, Ethylene Glycol, Perchloric acid, and permanganates.

·       Chlorine: With Ammonium, Acetylene, Butadiene, Benzene, and other petroleum fractions Hydrogen, sodium carbide, turpentine and finely divided metals.

·       Chromic Acid: With Acetic acid, naphthalene, camphor, alcohol, glycerol, turpentine and other flammable liquids.

·       Flammable liquids: With Ammonium titrate, chromic acid, hydrogen peroxide, nitric acid, sodium peroxide and halogens.

·       Potassium permanganate: With Glycerol, Ethylene, Glycol, Benazaldehyde and sulfuric acid.

·       Sulfuric Acid: With chlorates, per chlorates, permanganate and water.

v Toxic effects of chemicals

                          It is now well-known that some chemicals adversely affect the health of those hoop handle them or in hale their vapours. Apart from overt poisons, a number of chemicals are known to have various toxic effects. All the chemicals and reagent should be carefully handled. While handling chemicals, care to be taken so that they do not come in contact with skin, eyes, nose, etc. Adverse health effects of some laboratory chemicals are as folows.

TOXIC EFFECT OF CHLOROFORM

Ø  May cause Cancer.

Ø  May cause heritable genetic damage.

Ø  Harmful if swallowed.

Ø  Irritating to eyes, respiratory system and skin.

Ø  Harmful: Danger of serous damage to healthy by prolonged exposure through inhalation and if swallowed.

Ø  Harmful: Danger of serous damage to healthy by prolonged exposure through inhalation and if swallowed.

Chemical	Reported effects
	Acute	Chronic
Chloroform	Irritating to eyes, respiratory system and skin, DO NOT BREATHE VAPOR	May cause cancer.
Formaldehyde (Formalin)	Reparatory, skin, and mucus membrane irritation.	Pulmonary edema
Methanol	Necrosis, mucus membrane irritation.	Damage to retina and optic nerve
Acetone	Eye, Nose and throat irritation necrosis

SAFETY CONDITION:

      The following actions should be taken in the event of a spillage of dangerous chemicals.

·       Notify the safety officer and evacuate non-essential personnel from the area.

·       Attend to persons who may have been contaminated.

·       If the spilled material is flammable, extinguish all naked flames, turn of gas regulator to the room and adjacent areas and switch off electrical that may spark.

·       Avoid breathing vapor from spilled material.

·       Establish exhaust ventilation if it is sate to do so.

·       Secure the necessary items to clean up the spillage.

·       In the event of a large chemical spillage the room should be evacuated and windows opened if possible. If the material spilled is flammable, all naked flames in the room concerned and those adjacent should be extinguished and all electrical equipment that may spark switched off.

·        Spare cylinders should be stored in another building some distance from the lab.This store should be locked and identify by notice.

·       Compressed gas cylinders should be securely fixed (e.g. chained) to the wall or solid bench so that they are not dislogged by natural disasters.

·        Compressed gas cylinders and liquefied gas containers should not be placed near to radiators, Naked flames or other heat sources, or sparking   electrical equipment or direct sunlight

·        The main high pressure valve should be turned off when the equipment is not in use and when the room is unoccupied.

·       MSDS (Material safety data sheet) for the chemicals used in polio laboratory is available. It should be read carefully.

5.2 Laboratory Accidents-Electrical

Purpose: This SOP explains how to protect from electric hazards.

MATERIALS:   All electrical instruments, A.C. Biosafety cabinet lights, lights.

ELECTRIC HAZARDS: Electronic shock is life threatening, electrical faults may cause fires

SAFETY CONDITION:

·       Electrical units can cause electric shock and fire hazards. So when electricity failure occurs, first of all switches off all instruments and inform to electrician.

• Fire may occur when there is short circuit, then switch off instrument, MCB switch, see SOP of fire.
• If person is connected with electric current, remove with wooden stick, give first aid, and hospitalize the person.
• It is therefore essential that all electrical installation and equipments are inspected and tested regularly including earthing and are maintained by qualified electrician. Laboratory staff should not attempt to service any kind of electrical equipment.
• There should be protecting insulation to prevent fires, which protect people from electric shocked.
• All laboratory electrical equipment should be earthed.
• Avoid electrical wires cluttering.
• Remove uninsulated cables or electric wires.
• Laboratory staff should be made aware of the following hazards.

·       Wet or moist surfaces near electric equipment.

·       Long flexible electrical connecting cables.

·       Poor and perished insulation on cables.

·       Overloading of circuits by use of adapters.

·       Sparking equipments near to flammable substance and vapors.

·       Electrical equipment left switched on, but unattended.

5.3 Laboratory Accidents - Fire

Objective:  This procedure explains the procedures adopted to protect against fire in laboratory.

Purpose: This Sop explains how to protect if fire breaks in the laboratory.

GENERAL DESCRIPTION

The assistance of local fire prevention officers in the training of laboratory staff in fire prevention, immediate action in case of fire, and the use of fire-fighting equipment is desirable. Fire warning, instructions and escape routes should be displayed prominently in each room and in corridors and hallways.

COMMON CAUSES OF FIRE

• Electric overloading.
• Poor electrical maintenance
• Equipment left switched on unnecessarily.
• Overlong gas tubing.
• Naked flames.
• Deteriorated gas tubing
• Misuse of matches.
• Flammable and explosive chemicals stored in ordinary refrigerators.