Betriebsanweisung Genlabore

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Dec 11, 2012 (4 years and 6 months ago)

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Abteilung

für

am Universitätsklinikums Bonn

Stand:
09
.
10
.2007








SAFETY INSTRUCTIONS


ACCORDING TO §12
GENTECHNIK
-
SICHERHEITSVERORDNUN
G

FOR LABORATORIES OF
BIO
SAFETY LEVEL 1

AND

§12 (1)
BIOSTOFFVO



Unbefugten

Zutritt verboten
















Gentechnischer
Arbeitsbereich


S1


Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
2







MS UKB


Inhaltsverzeichnis



1.
Area of application

2. Responsible
persons


3. Genetic engineering work

4.
Potential hazards from

GMOs

5.
Facility work practices


5.1 A
c
cess to the laboratory


5.2 General instructions


5.3
Personal and laboratory Hygiene


5.4
Interdictions


5.5 Protective Equipment

6.
Emergency management


6.1
Bio
logical spills


6.2 Fire


6.3. Leakages

7. First Aid

8.
Waste handling

9.
References and Guidelines

10.
S
pecific regulations











Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
3







MS UKB


1.
Area of Application

These Instructions are to be followed in the
biosafety level 1
laboratory (
Genlabor,
(
Reg.
-
No.
53
.02.01
-
K
-
1.
x
x/
xx
)
) in the

______________________

des Universitätsklinikums Bonn,
Sigmund
-
Freud
-
Str. 25, 53127 Bonn.


The labortatory comprises

the following rooms:


Laboratory rooms
: (
x
x
xx

/

x
x
xx

/
x
x
xx

)

Scullery / Autoklave

(
x
x
xx
)

Cooling chambers
: (
x
x
x
x
)

Corridor
: (
x
x
xx
)


All Doors leading into the laboratory are

labeled as "Genetic Laboratory Safety Level 1"
("
Genlabor Sicherheitsstufe

1")
.


The rooms
x
x
xx

/
x
x
xx

are to be used as common rooms.


2. Responsible P
ersons


Head of Laboratory

(
Projektleit
er
)


x
x
xx

0228
xxxxxxxx

x
x
xx

0228
xxxxxxxx

x
x
xx

0228
xxxxxxxx



Biosafety Officer

(
Beauftragter für biologische Sicherheit
)


x
x
xx

0228
xxxxxxxx



Responsible for

operational safety



Dipl.
-
Ing
.

B. Bergmann

0228
287
15088



Company medical Offic
er

(Betriebsarzt)


Dr.
Packbier

88 16176



In case of poisoning

(Vergiftungszentrale)


Univ. Kinderklinik, Adenauerallee 119, 53113 Bonn

88
-
132 11



Technical
emergency
Service

0228 287
-
9



Fire brigade/Emergency medical services:

0
-
112

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
4







MS UKB


3. Geneti
c engineering work


All projects performed in this laboratory are of biosafety level 1
. No projects of higher
biosafety levels according to
§12
GENTECHNIK
-
SICHERHEITSVERORDNUN
G

are
allowed.
Additionally in rooms
x
x
xx
,
x
x
xx

and
x
x
xx

projects of biosafety le
vel 2 according to
§12 (1)
BIOSTOFFVO

are allowed. Specific biosafety instructions are available for these
rooms.


The genetic engineering work includes the production,
use,
storage, transportation
,
disinfection

and disposal of the genetically modified org
anisms.


The head of laboratory has to be informed in any unusual event.


4.
Potential hazards from

GMO
s


The genetically modified organisms
used in this laboratory
belong to risk group 1. This
means that their appropriate usage
-

according to these instru
ctions
-

does not endanger
healthy people or the environment.
In case of immunospupression, pregnancy or allergies
special rules of conduct have to be applied.
A comprehensive risk assessment
has do be
done before starting of biosafety level 1 projects and

must accompany the filed records of the
genetic engineering work

(§§ 6,7 GenTG, GenTSV Supplement 2)
.




5.
Facility work practices


Al
l projects have to be performed according to

the general rules of microbiological
techniques and genetic engineering saf
ety instructions

(Gentechnische
Sicherheitsverordnung, GenTSV)
.


5.1 Access to the genetic laboratory



Pregnant and nursing woman are not allowed to work in a laboratory performing
projects comprising pathogens for humans.



Visitors or maintenance personnel

have to be accompanied by instructed laboratory
staff.



All persons entering the laboratory have to wear lab
coats
, protective equipment and
appropriate footwear

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
5







MS UKB




All laboratory staff
has to be informed about the safety instructions before starting to
work

and afterwards in yearly intervals. The safety instructions have to comprise
information about this safety manual, hazardous goods and other potential dangers in
the laboratory. All participants have to sign their attendance.



Cleaning personnel has to be
instructed in yearly intervals according to this safety
instructions. All participants have to sign their attendance.



Staff which is inexperienced in microbiology, virology or cell biology have to be
trained and accompanied by an experienced person.


5.2 G
eneral instructions



The laboratory has to be labeled with signs for
“Biosafety Laborators level 1”
(
Gen
technischer Arbeitsbereich

S1
) and “Working personnel only”.







All laboratory staff has to be familiar with location and func
tion of disinfectants, eye
and body showers, first aid kits and fire extinguishers

as well as procedures for
evacuation in case of emergency.




All laboratory staff has to wear appropriate protective equipment (e.g. gloves,
protection glasses) when working
with hazardous chemicals. When working with UV
light a protective shield is necessary. Avoid direct exposure to eyes and skin.



All laboratory rooms have to be clean and tidy. Spare material has to be stored in the
storage rooms and only equipment and goods

of daily usage should be kept on the
benches.



No office work, despite recording of the experiments, should be done in the
laboratory. Manuals and catalogues have to be stored in documentation rooms.



Door and windows have to be closed during genetic engine
ering work. It is allowed to
open windows if no genetic engineering work is done and if the airflow of the safety
cabinets is not disturbed.



Usage of pipetting aids is mandatory. Pipetting by mouth is prohibited.



Syringes, Scalpels, sharp disposable knifes

etc should be used only if necessary.
They have to be disposed in special waste containers. They have to be autoclaved
before disposal.

Gentechnischer
Arbeitsbereich


S1

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
6







MS UKB




The Formation of aerosols has to be avoided. Beware of the formation of aerosols
especially when executing the followin
g procedures: Shaking, stirring, decanting,
pipetting, centrifuging, working with ultrasound and inoculating.

The following strategies are helpful to avoid aerosols:

o

Usage of closed bottles or containers

o

Avoidance of bubbles

o

Slow decanting or pipetting di
rectly to the walls of the destination bottles.

o

Usage of safety cabinets



The identity of the used organisms and GMOs has to be checked regularly This can
be done by the following procedures:

o

Cultivation on special plates with minimal essential ingredients

o

Cultivation with antibiotics

o

PCR, restriction analysis or sequencing



In addition to this safety instruction every member of the laboratory has to pay
attention to the special manuals from the manufacturer of the devices used in the
laboratory. They are sto
red beside the equipment or in room 404
-
1.



When transporting GMOs closed, unbreakable, aerosol and liquid safe containers
have to be used.



GMO
s

are to be stored in a appropriate manner, e.g. in screwed tubes in the
freezers.


5.3
Personal and laboratory H
ygiene

The instructions provided in the “disinfection manual” (Hygieneplan S1) are part of
this safety instruction. All staff has to pay attention to them.




After finishing the laboratory work all benches have to be cleaned with the
appropriate disinfectan
ts stated in the “disinfection manual”.



70% Ethanol is not allowed for disinfection of large areas due to possible
inflammation. Ethanol should only be used if no other disinfectant is suitable. The
usage of ethanol sprayers should be avoided if possible.



Before leaving the laboratory area hands should be disinfected and cleaned
according to the “disinfection manual”.

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
7







MS UKB




Equipment has to be cleaned regularly:

o

Centrifuges have to be cleaned directly after contamination or at least once a
month.

o

The safety cab
inets have to be cleaned directly after use or at least every two
month.

o

Incubators have to be cleaned at least every two month. Before cleaning they
have to be unplugged and cooled down / heated up to room temperature.

o

Containers with disinfection solutio
ns have to be cleaned at least once a weak
and refilled with fresh disinfection solution.



When cleaning electric devices or devices with open fire or hot surfaces special
attention is required. Be sure to unplug electric devices first when possible.



Vermin

has to be destroyed by professionals.


5.4
Interdictions



Eating, drinking and smoking is prohibited in the laboratory.



Storage of food and luxury food as well as cosmetics is not allowed inside the
laboratory. They can be stored in room 208 (common room)
.



It is not allowed to enter the common rooms with protective equipment.



Pipetting with the mouth is prohibited. The usage of pipetting aids is mandatory.



The usage of vacuum pumps is not allowed if not sterile filter is used to prevent
contamination of th
e air with GMOs.



It is prohibited to store GMOs and other material in the corridors.


5.5 Protective equipment



All persons have to wear lab coats inside the laboratory. The lab coats have to be
cleaned at least once a weak. This will be centrally done by t
he UKB laundry.



Lab coat
s and protective equipment have to be dropped of before leaving the
laboratory.



Disposable gloves have to be disposed correctly. If contaminated with GMOs they
have to be autoclaved first.



Lab coats and normal clothing have to be st
ored separately. The normal clothing can
be stored at the writing desks.

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
8







MS UKB


6.

Emergency management



Keep calm, don’t panic!



Warn endangered people and, if necessary, evacuate rooms.



End dangerous and endangered experiments and turn of gas, electricity (by pr
essing
the red buttons besides each door) and water (below the sink) if necessary. Keep in
mind that cooling could still be necessary!



Inform the head of laboratory in any case of emergency!


6.1
Biological spills

Areas, where biological material is spill
ed, must be
labeled

immediately.


If material containing GMOs is spilled it has to be inactivated immediately according to the
following disinfection strategies:



Surfaces

Wear protective gloves!

Absorb spilled material with paper tissue and dispose the ti
ssue into autoclavable
bags. Disinfect the surface according to the “disinfection manual”. If glass is broken
disinfect the hole area and remove the glass pieces afterwards!



Devices

Wear protective gloves!

Absorb spilled material with paper tissue and disp
ose the tissue into autoclavable
bags. Disinfect the device according to the “disinfection manual”.

When cleaning electric devices or devices with open fire or hot surfaces special
attention is required. Be sure to unplug electric devices first.



Clothing

D
rop of the contaminated clothing and disinfect or autoclave it.



Skin

Disinfect the contaminated skin according to the “disinfection manual” and wash of
the disinfection solution afterwards.



Eyes

Water the eyes with the eye shower for at least 5 minutes. S
peak to the company
medical officer afterwards.



Mucosa

Water the contaminated area for at least 5 minutes. Speak to the company medical
officer afterwards

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
9







MS UKB


If you suffer injury from a contamination speak to the emergency unit of the surgical
department
or
the ophthalmic department immediately. If you are at home contact the next
hospital immediately.

Emergency surgery: 88
-
15109 / 152 31

Emergency unit of the Ophthalmic clinic: 88 15505 / 156 39


Any case of injury has to be reported to the head of laborator
y. He has to report the injury to
the Department of operational Safety using the “Unfallbericht” form.



6.2 Fire

To blot out small fires use the fire extinguishers installed in the laboratory.

In any other case follow the fire emergency plan.


Fire brigad
e: 0
-
112



6.3 Leakage

In case of a leakage disinfect the contaminated area according to the “disinfection manual”.
To avoid leakage of centrifuges do not overfill tubes!


7. First aid



First aid kits can be found in the corridor netx to rooms
x
x
xx

and
xxxx
.

The following persons are trained in first aid:

xxxxxxx

xxxxxxx

xxxxxxx




Injuries

o

Apply first aid if possible

o

If necessary call an ambulance. Inform the medical personnel about the
possibility of infections though biological material or hazardous goods.

o

Injuries have to be reported to the head of laboratory immediately.

o

If you suffer from an injury later on speak to the emergency unit of the surgical
department or the ophthalmic department immediately. If you are at home
contact the next hospital immediat
ely.

o

Information about injuries in combination with GMOs have to be recorded and
stored for 10 years.

Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
10







MS UKB




Inhalation or
swallowing

of GMOs or hazardous goods

o

Inform the head of laboratory immediately.

o

Speak to the companies medical officer immediately if treat
ment is necessary.
It is important to provide information about the inhal
a
ted or swallowed
material.

o

If necessary call an ambulance and apply first aid.



8.
Waste handling

All waste contaminated with GMOs has to be auto
c
laved.

The Autoclaved is to be used

by trained personnel only.

The autoclaving has to be done at 121°C, 1 bar for 20 minutes. The autoclave can be found
in room
x
x
xx
.

The autoclave has to be checked at least once a year. Bioindicator tests are available from
the Hygiene department. The res
ults of the tes
ts

ha
ve

to be documented.

After autoclaving the waste can be disposed in a normal way.


9.
References and guidelines


Laws / Regulations

GenTG

Gen
technikgesetz

ZKBS

Verordnung über die Zentrale Kommission für die
Biologische Sicherheit

GenTAufzV

Gentechnik
-
Aufzeichnungsverordnung

GenTSV

Gentechnik
-
Sicherheitsverordnung

GenTAnhV

Gentechnik
-
Anhörungsverordnung

GenTVfV

Gentechnik
-
Verfahrensverordnung

GenTBetV

Gentechnik
-
Beteiligungsverordnung

GenTNotfV

Gentechnik
-
Notfallverordnung

BiostoffV

Biostoffverordnung

IfSG

Infektionsschutzgesetz

GefStoffV

Gefahrstoffverordnung



GuidelinesTechnische Regeln

TRBA

Technische Regeln für Biologische Arbeitsstoffe

TRBA Reihen:

100, 200, 300, 400, 500


Further information can be obtained for the department of operational safety or the related
homepage.


Safety indtructions

for

Laboratories of
bios
afety level 1

Ab
teilung
für , UK Bonn

page
11







MS UKB


10.
Specific Regulations




Duty of notification

The
Head of laboratory

must be informed about all incidents, w
hich are unusual for
genetic engineering work.



Instructions

All laboratory staff has to be informed about the safety instructions before starting to
work and afterwards in yearly intervals. The safety instructions have to comprise
information about this sa
fety manual, hazardous goods and other potential dangers in
the laboratory. All participants have to sign their attendance.



Duty of recording

Only
b
iosafety level 1 projects are permitted in this laboratory.
All genetic engineering
work has to be recorded
using the templates provided by the head of laboratory. The
form provides information on the transferred gene, the donor and receiver organism,
the vector and the GMO itself. All this information have to be stored for 10 years after
the final experiment of

the project.



Fines

Contravention of the regulations
stated in the genetic enginee
ring laws can
result in fines up to 50.000€ and imprisoning for up to 5 years. In addition
indemnification up to 85.000.000€ can be accrue.



Bonn,







Head of Laboratory


B
SO