rDNA/IBC Protocol Form - Office of Research and Sponsored Projects

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Dec 14, 2013 (3 years and 7 months ago)

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University of Texas at El Paso

S
ynopsis for Research Involving the Use of

Infectious Agents, Toxins or recombinant DNA
(rDNA)



The following
guidance is required

for
preparing

research
proposals and other plans

involving
recombinant DNA, infectious age
nts or toxins

for review
by the Institutional rDNA/Biosafety
Committee (IBC)
All protocol information must be typed.
The
NIH Guidelines

can be found
at:

http://oba.od.nih.gov/rdna/nih_guidel
ines_oba.html
. The Synopsis is submitted to the Office of
Research and Sponsored Projects (ORSP), IBC Coordinator
.


Protocol

Submission Process
:
T
h
e

IBC Coordinator will administratively screen all protocols.

The
IBC Chair

will assign an IBC committee m
ember
an
d the Biosafety

Officer (BSO)
to

provide
the committee with a summary

at
the next

IBC
committee meeting.

All protocols must be
reviewed at full IBC meeting. After review and discussion of each protocol, the IBC members
will
vote to
table, approve
or approve with modifications
. All approved protocols will be active for
a
three year term and subject to the
NIH Guidelines for Research Involving Recombinant DNA

Molecules

(see

http://oba.o
d.nih.gov/rdna/nih_guidelines_oba.html
). Compliance with the NIH
Guidelines is a requirement for all research performed at the University
.


Progress reports will be required annually from PIs who wish to keep their IBC protocol
s
active.
Progress report

should provide sufficient information to enable the IBC to make an
informed risk assessment of the project.

Any
spills

or injuries that occur while conducting
research covered by IBC protocols must be reported to the Biosafety

Officer

(747
-
7179)
and the
IBC
Chair (747
-
8715) or IBC
Coordinator (747
-
7007) immediately.

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I.

General Information
:


P
rincipal
I
nvestigator
:







D
ept
:








Funding Agency
:







Funding
P
eriod
:








Protocol Title
:







Attach a summary in non
-
technical lay terms of the overall aim and scope of the research
protocol.

The summary s
hould not be more than a page in length.








II.

Biological Agents Used:

List and describe organism(s) used such as bacteria, parasites, toxins, fungi, viruses, cell
lines, and animals to be used. Include specific name, strain, source infor
mation, host(s) and
included bioengineered controls, if any. Also, describe the biological agent or microorganism
characteristics (e.g. virulence, pathogenicity, environmental stability and infectious dose)








What is the biological safe
ty level (BSL) for this project:


BSL1


BSL2


BSL3


See Biosafety in Microbiological and Biomedical Laboratories (BMBL)

5
th

Edition at:
http://www.cdc.gov/biosafety/publications/bmbl5/index.htm


III.

Protocol Classification

Does your protocol involve the generation of transg
enic
organisms,
animals or recombinant
DNA?



Yes
, If
YES
,

complete the entire form and all

sections.



No
, If
NO
, you can skip Section IV and Section V of the form.


IV.

Recombinant DNA Classification
:

(Check 'yes' or
'no' to the following questions
.

The relevant section of the
NIH Guidelines

(see

http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
)

is referenced for each question.


1.

Does your project include
deliberate transfer of a drug resistance
trait to pathogenic microorganisms that are not known to acquire
the t
rait naturally (
Section III
-
A
-
1
-
a
, experiments falling under this
section of the NIH Guidelines require IBC approval, Recombinant
DNA Advisory (RAC) Committee review and NIH Director
Approval before initiation. For example
,

the introduction of the
gene en
coding chloramphenicol resistance into
Rickettsia
conorii
)?


Yes


No




2.

Does your project include cloning toxin molecules with a

lethal dose

(
LD
50
)

of less than 100 nanograms per kilogram body weight
(Section III
-
B
-
1,
e
xperiments falling under this section of the NIH
Guidelines require IBC approval, and submission to the NIH Office
of Biotechnology Activities [OBA] before initiation. For example the
cloning of the gene coding for the botulinum toxin.
)?


Yes


No

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3.

Does your project include experiments involving the deliberate
transfer of recombinant DNA, or DNA or RNA derived from
recombinant DNA, into one or more human research participants
(Section III
-
C
-
1,
experiments falling under
this section of the NIH
Guidelines require IBC approval,

Recombinant DNA Advisory
(RAC) Committee review

and
IRB approval
before initiation
)?


Yes


No




4.

Does your project include experiments using Risk Group 2, Risk
Gro
up 3, Risk Group 4, or Restricted Agents as host
-
vector
systems (Section III
-
D
-
1)

that would require BSL
-
2 or BSL
-
3
containment

(
experiments falling under this section of the NIH
Guidelines require IBC approval

before initiation )
?


Yes


No




5.

Does your project include experiments in which DNA from Risk
Group 2, Risk Group 3, Risk Group 4, or Restricted Agents is
cloned into nonpathogenic prokaryotic or lower eukaryotic host
-
vector systems (Section III
-
D
-
2
,
experiments fa
lling under this
section of the NIH Guidelines require IBC approval
)?


If Yes, is the cloning into an E. coli K
-
12 strain or K
-
12 derivative

(
If
Yes, this work falls under Section III
-
F
-
6 and Appendix C
-
II
)
?


Yes









Yes


No





No


6.

Does your project include experiments involving the use of infectious
DNA or RNA viruses or defective DNA or RNA viruses in

the
presence of helper virus in tissue culture systems (Section III
-
D
-
3
,
experiments falling under this section of the NIH Guidelines require
IBC approval
)?


Yes


No




7.

Does your project include experiments involving wh
ole animals in
which the animal’s genome has been altered by

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-
a
-
㐬 III
-
b
-
3



If
only Section III
-
E
-
3 is applicable then the
se experiments may be
initiated at the same time as IBC registration is in process
)?


Yes


No




8.

Does your project include experiments involving whole plants
(Section III
-
D
-
5, III
-
E
-
2
,
If only Section III
-
E
-
2

is applicab
le then
these experiments may be initiated at the same time as IBC
registration is in process
)?


Yes


No




9.

Does your project include experiments involving more than 6 liters of
culture (Section III
-
D
-
6)?


Yes


No




10.

Does your project include experiments involving the formation of
recombinant DNA molecules containing two
-
thirds or more of the
genome of any eukaryotic virus (Section III
-
E
-
1
,
If only Section III
-
E
-
1

is applicable then the
se experiments may be initiated at the same
time as IBC registration is in process
)?



Yes


No




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V.

Description of rDNA Experiments

Describe

the specific
methods and
experiments that will be performed under the rDNA
pr
otocol and include:


1.

List IRB, IACUC or IBC protocol number as references, where applicable;








2.

Provide literature references, if appropriate. (For example,
Naldini, L., Blomer, U., Gage,
F. H., Trono, D., and Verma, I. M. (1996) Efficient Transfer, Integration, and

Sustained
Long
-
Term Expression of the Transgene in Adult Rat Brains Injected

with a Lentiviral

Vector. Proc. Natl. Acad. Sci. USA
93
, 11382
-
11388
);








3.

Include methods or processes which will generate hazardous aerosols such as sonicating
or cell sorting (For example


In addition to the experiments and methods de
tailed above,
we will be extracting proteins via sonication. Sonication methods include … Additionally,
hearing protecting muffs or ear plugs will be provided to lab personnel doing sonication).








4.

Describe the m
anipulations that will b
e performed under the protocol. For example, (a)
describe the cloning of gene Y into bacterial plasmid for protein expression; (b). describe
the cloning of protein X into a lentiviral vector; (c) describe the experiments leading to the
cloning of transgen
e Z for generation of transgenic animals:








5.

What is the source of rDNA, DNA, RNA to be inserted or cloned


include species of
organism from which it is derived.








6.

What is the nature of rDNA, DNA, RNA to be inserted or cloned. For example is it a
structural gene or oncoge
ne.








7.

What is the host system to be used? For example, Rat cardiomyocytes and HEK
-
293
cells.








8.

What is the vector(s) to be used? Include information such as product literature, or vector
map describing construction o
f vector. (For example, Lentiviral vector, Invitrogen
ViraPower
TM
)
:








9.

Are any helper virus or packaging cells used? (For example 293FT cell line);









VI.

Location of Research:

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Please list all locations where work will b
e conducted. Include building and room numbers for
storage areas, work areas, biosafety cabinet locations and autoclave locations. (For
example, stocks and materials will be stored in room VRB 6.218, we will dedicate the BSC in
room VRB 7.456 for lentivi
ral work and the BSC in room 7.268 for all other work, animal
studies will be conducted in Research Hall 9.236 and finally the biowaste will be
decontaminated using the autoclave in room 7.345):








VII.

Indicate the Personal Protective Equipment that will be used to perform the work:



Single
-
use disposable latex gloves



Single
-
use disposable nitrile gloves


Safety glasses





Safety goggles


Open
-
front Laboratory coat




Closed
-
front lab gown


Dust mask






N
-
95 Filtering Face piece


Sleeve guards






Face shield


Ear plugs






Ear muffs


Shoe covers






Hair bonnet


2 pairs of single
-
use disposable gloves




VIII.

Biological Safety Practices:


1.

Provide a description of the appropriate

containment conditions and biosafety practices
that will be implemented for the proposed project based on risk assessment and biosafety
level. Certain experiments have special considerations that must be taken into account
such as the use of cell sorters

for infectious agents or the use of viral vectors. See,
CDC/NIH Publication, Biosafety in Microbiological and Biomedical Laboratories (BMBL)
5
th

Edition:;

http://www.cdc.gov/biosafety
/publications/bmbl5/index.htm
.








2.

List the disinfectant(s) to be used under this protocol?







3.

Describe biological waste disposal methods:







4.

Will Biosafety cabinets be used? If yes, include last certi
fication date:







5.

Do you intend to ship infectious substances or hazardous chemicals?

Yes


No
.

If
YES
, please, contact Environmental Health and Safety (EH
&
S

x7124
)

to make
arrangements.


6.

List the 24 hour

emergency contact phone number for the PI, lab director or responsible
person to contact.








IX.

Protocols Involving Lentiviral Vectors

must describe the criteria for risk assessment of
the lentivirus vector(s) used
:

YES

N
O


See also, the NIH guidance document, Biosafety Considerations for Research with Lentiviral
Vectors,

http://oba.od.nih.gov/rdna_rac/rac_guidance_lentivirus.html


1.

Describe
the nature of the vector system and the potential for regeneration of replication
competent v
irus from the vector components.







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2.

Describe
the nature of the transgene insert (e.g., known oncogenes or genes with high
oncogenic potential may merit special care)
.







3.

Describe the anticipated
vector titer and
anticipated
total amount of vector

to be produced.







4.

Describe
the inherent biological containment of the animal host, if relevant.











X.

Personnel:

Please list the names of the individuals who are covered under this protocol.
Include their full name, title (faculty, post

doctoral, graduate and undergraduate students,
visiting scientist
, and staff
), UTEP 800 ID #, and laboratory experience directly related to the
experiments covered under the protocol, UTEP Safety Classes and date taken.

All
individuals must have current
safety training in order to work under the protocol. Please
contact EHS for training dates, if necessary.

Please be informed that if all individuals are
not current with all training requirement
s
, this protocol will not be reviewed.


Name

Title

UTEP
ID

Ex
perience

Required
Safety Class
es


Date
Taken

Principal
Investigator:







PI/




















Basic Laboratory Safety








Blood
b
orne Pathog
ens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory S
afety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program
































Basic Laboratory Safety








Blood
b
orne Pathogens








Occupational Health Program







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XI.

Inves
tigator Agreement:


I attest that t
he information
provided or attached
is accurate and complete. I am familiar with
and agree to abide by provisions of the current
NIH Guidelines for Research Involving
Recombinant DNA

Molecules

and accept the responsibili
ties listed in Section IV
-
B
-
7
.


As the Principal Investigator, I accept responsibility for making sure all laboratory
p
ers
onnel

involved in the project have been appropriately trained. All research personnel
are
familiar
with and understand the potential b
iohazards and relevant biosafety practices, techniques,
and emergency procedures

associated with this research protocol as dictated by the CDC
and NIH document Biosafety in Microbiological and Biomedical Laboratories, 5thd Edition
(
http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
).

I certify that I will immediately
report a
ny injuries
or spills
that occur while conducting research covered by
this
IBC
protocol to the UTEP Biosafety
Office
r

(747
-
7179) and the IBC C
hair (747
-
8715) or IBC
C
oordinator (747
-
7007
)
.



S
ignature
:

D
ate
:









Department Chair:

_________________________________________
_ Date:










IBC REVIEW RESULTS



Date of Full Committee Mee
ting:








IBC Committee Determined Containment Level:








Applicable NIH Guidelines Section:







Protocol Status:


Tabled

N
eeds Modification (not approved)



Approved




Conditionally Approved with Listed Modifications


Modifications and
Remarks:







IBC Chairman:


Date:








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STANDARD OPERATING PROCEDURES FOR HANDLING INJURIES, BIOLOGICAL SPILLS AND
BIOHAZARDOUS WAST
E IN THE LABORATORY



Purpose:

This Standard Operating Procedure outlines the necessary steps to take when someone is injured,
when cleaning up biological spills or handling biohazardous waste in Dr.





laboratory.


Scope:

Everyone working
in Dr.






laboratory who handles human cell lines, human blood,
microbiological cultures, parasites, viruses and other biological material such as recombinant DNA
or proteins.


NOTE: This SOP must be posted in the lab and be distributed to

all current and incoming
research students and staff.


Procedures:

1.

Injury to an Individual in the Lab (i.e. needle stick, cut, biological/chemical exposure
incident


splash, etc.):

a.

Immediately stop work and flush affected area with soap and water for 15
minutes.

b.

If the injury is a Medical Emergency call 911 or campus police x5611.

c.

Secure all infectious materials.

d.

Notify Dr.






at (



)






and EH&S Biosafety Officer at (915) 726
-
7887 or
Campus police x5611 after working hours. This is very important as the University
maintains an ongoing log/list of spills and injuries and as applicable repo
rts these
as required under the
NIH Guidelines for Research Involving Recombinant DNA
Molecules
.

e.

Use First Aid Kit located in the Glass Wash Room or Tissue Culture Room
whichever is the closest.

f.

If during working hours seek medical attention at the Student

Health Center
under the UTEP Occupational Health Program.

g.

Dr.






will complete the Workers Compensation Injury/Incident Report form
documenting the route of exposure and the circumstances under which the
incident occurred.


2.

Small Spill Decontamination and Clean Up (less than 1 liter):

a.

Stop work and secure all ite
ms you are working with.

b.

Replace any contaminated personal protective equipment (PPE).

c.

Make sure you are wearing the appropriate PPE such as disposable gloves, lab
coat and eye protection (safety glasses or goggles).

d.

Using the Lab Spill Kit provided by Env
ironmental Health & Safety, take the
absorbent pads and place over the spill area.

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e.

Make a fresh solution of 10% bleach and pour over the absorbent pads. The
absorbent pads absorb the spill, help contain the 10% bleach solution and help
prevent splattering
.

f.

Let the bleach solution inactivate and decontaminate the biological material for a
minimum of 15 minutes.

g.

Report the spill to Dr.






at (



)




and EH&S Biosafety Officer (915) 726
-
7887 or after hours at x561
1. This is very important as the University maintains
an ongoing log/list of spills and injuries and as applicable reports these as
required under the
NIH Guidelines for Research Involving Recombinant DNA
Molecules
.

h.

Soak up and clean up the excess bleach
solution and decontaminated material
with extra absorbent pads or paper towels.

i.

Dispose in the red biowaste can.


3.

Large Spill (greater than a liter):

a.

Stop work immediately, secure all items and avoid inhaling airborne aerosols.

b.

Notify others to leave room
immediately.

c.

Label the area off
-
limits for at least 30 minutes. This allows the ventilation
system to purge the air.

d.

Remove contaminated PPE and or clothing, turn exposed clothing inward, and put
in autoclave bag or red biohazard bag. Wash all exposed sk
in with soap and
water.

e.

Report the spill to Dr.






at (



)






and EH&S Biosafety Officer (915) 726
-
7887 or after hours at x5611 Campus Police. This is very important as the
University maintains an ongoing log/list of spills and injuries and as applicable
reports these

as required under the
NIH Guidelines for Research Involving
Recombinant DNA Molecules
.

f.

After at least 30 minutes, EH&S personnel will enter the area to clean up the spill.


4.

Liquid Biohazardous Waste Disposal:

All liquid biological waste from the lab must
be treated prior to disposal. Examples of
biological waste include cell lines, recombinant DNA, recombinant proteins, and bacterial
cells. The procedures below outline the steps to take to treat liquid biohazardous waste
generated in Dr.






lab:

a.

Always wear appropriate PPE such as disposable gloves, lab coat and eye
protection (safety glasses or goggles) when working with biohazardous waste.

b.

When liquid biohazardous waste is anticipated to be generated, add 100 ml of
undiluted Clorox B
leach into a 1 L beaker.

c.

Label beaker appropriately as to its contents

d.

If more than 1L of liquid biohazardous waste is anticipated prepare a second 1 L
beaker by adding 100 ml of undiluted Clorox bleach to the second 1L beaker.

e.

As experiments are performed

and completed pour the biological waste into the
beaker with the Clorox Bleach.

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f.

Once experiments are complete and if the beaker is less than 1L add water to
bring the volume to 1L.

g.

Once the beaker is full the Clorox bleach has been diluted to a 10% soluti
on.

h.

Let 10% bleach and biological waste solution stand for at least 1 hour.

i.

Dispose of the solution with care to avoid splatter down the lab sink and rinse
beaker.


4.

Solid Biohazardous Waste Disposal:

The procedures below outline the steps to take to treat
solid biohazardous waste
generated in Dr.






lab:

a.

All solid lab waste that has come in contact with biological waste from the lab
must be treated prior to disposal. Examples of biological waste include used
personal protective equipment such as disposable gloves, paper towels, pipette
tips, d
isposable Petri dishes, pipettes and culture flasks.

b.

Always wear appropriate PPE such as disposable gloves, lab coat and eye
protection (safety glasses or goggles) when working with biohazardous waste.

c.

Place all potentially contaminated or contaminated ite
ms in a red biohazardous
waste bag.

d.

Once the bag is ¾ full close bag and place autoclave tape on the bag.

e.

Take the biowaste bag to the Glass Wash Room and place in the autoclave.

f.

Complete autoclave log book entry and autoclave biohazardous waste at least 3
0
minutes following manufacturers’ recommendations for autoclave operation.

g.

Once the autoclave cycle is complete the load within has been sterilized if the
autoclave tape has turned color and the autoclave display shows no errors.

h.

Place the red autoclaved
biohazardous bag into a large 55 gallon red bin for
removal from the facility by EH&S.

i.

If the autoclave tape did not turn color and or autoclave display indicates errors
occurred during operation or an incomplete cycle the load has not been sterilized.

j.

Immediately call the Bioscience Building Manager at x6881 and EH&S Biosafety
Officer (915) 726
-
7887 or x7179 as the load is still considered biohazardous.


Other special concerns, if applicable: