200292
NucliSENS® Lysis Buffer
For in vitro diagnostic use.
INTENDED USE
®
NucliSENS Lysis Buffer is intended to be used for the release of total nucleic acid from biological specimens.
SUMMARY AND EXPLANATION
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Biological samples prepared using NucliSENS Lysis Buffer are suitable for use in nucleic isolation procedures based on
(1-9)
. The lysis procedure described below has been designed as a generic laboratory protocol.
Boom chemistry
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bioMérieux has performed validation for a limited number of human specimen types using NucliSENS Magnetic
®
Extraction Reagents and the NucliSENS miniMAG.
PRINCIPLE OF THE METHOD
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Biological specimen is added to NucliSENS Lysis Buffer, which contains guanidine thiocyanate. Any viral particles or
cells in the specimen will be disrupted, releasing all nucleic acids that may be present. RNases and DNases in the
(1).
specimen will be inactivated
REAGENTS
For in vitro diagnostic use.
Contents
Catalogue number: 200292.
®
NucliSENS Lysis Buffer (2 ml) contains sufficient reagents for up to 48 isolations.
Contents
Component
Description
Lysis Buffer
LYS
48 x 2 ml
Contains 50 % guanidine thiocyanate*,
<2 % Triton X-100, <1 % EDTA
1 package insert downloadable from www.biomerieux.com/techlib.
Warnings and Precautions
* Signal Word: WARNING
Hazard statement
EUH031: Contact with acids liberates toxic gas.
H302 + H312 +H332: Harmful if swallowed, in contact with skin or if inhaled.
H412: Harmful to aquatic life with long lasting effects.
Precautionary statement
P261: Avoid breathing dust/fume/gas/mist/vapours/spray.
P273: Avoid release to the environment.
P280: Wear protective gloves/protective clothing/eye protection/face protection.
P301 + P330 + P331: IF SWALLOWED: Rinse mouth. Do NOT induce vomiting.
P302 + P352: IF ON SKIN: Wash with plenty of water.
For further information, refer to the Material Safety Data Sheet
o
o
o
Certain reagents contain guanidine thiocyanate. Refer to the hazard statements "H" and the precautionary
statements "P" above.
Warning: Buffers containing guanidine thiocyanate should not be mixed with cleaning solutions containing
bleach. Liquid waste from extraction and isolation procedures containing guanidine thiocyanate must not be
mixed with other laboratory waste. This will prevent potentially harmful chemical reactions from occurring.
Reagent spillages should always be dealt with appropriately (see Procedural Precautions).
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Reagent Storage
o
o
o
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o
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Store at ambient temperature 2 to 30 °C.
Protect reagents from excess heat or light.
Reagent storage in the area of the laboratory dedicated to the isolation of nucleic acids is recommended.
Only the required quantities of reagents should be removed from storage.
Return any unopened reagents to storage immediately.
Storage of opened reagents is not recommended.
Reagent Stability
o
o
o
o
o
o
Reagent stability after opening cannot be guaranteed. Opened reagents should not be stored for later use.
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NucliSENS Lysis Buffer may have a slight to appreciable yellowish color. This coloration is normal and will not
have any effect on the performance of the reagents.
Expiry dates shown on kit and component labels indicate the date beyond which reagents should no longer be
used.
®
Storage of NucliSENS Lysis Buffer at 2 to 8 °C may give rise to the appearance of crystals due to the high salt
concentration. These crystals will dissolve during reagent preparation.
Other changes in the physical appearance of reagents may indicate instability or deterioration and they should
not be used.
Please contact your local bioMérieux representative for assistance if you are in doubt about the suitability of
reagents for use.
SPECIMEN
Specimen Type and Collection
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NucliSENS Lysis Buffer is intended to be used for the extraction of nucleic acids from a wide range of biological
specimens. For examples, see references 2 to 14.
o Consult individual instructions for use for further details when using these reagents in conjunction with other
bioMérieux products.
Specimen Stability
o
Consult individual instructions for use for further details when using these reagents in conjunction with other
bioMérieux products.
PROCEDURE
o
Room temperature = 15 to 30 °C.
Procedural Precautions
o
Store and prepare reagents for nucleic acid release in the laboratory area where nucleic acid isolation is to be
performed.
Avoid contamination or specimen-to-specimen carry-over:
o Perform nucleic acid isolation in a dedicated laboratory area (preferably a self-contained area or a fume hood)
that is not connected to laboratory areas where amplification or detection is to take place.
o Keep all tubes and vials closed when not in use.
o Pipettes and other equipment that have been used for nucleic acid isolation must not be used in laboratory
areas where nucleic acid amplification or detection is to be performed.
o Use a fresh pipette or pipette tip for each pipetting action.
o Use pipettes with aerosol resistant tips for fluids possibly containing nucleic acid.
o Only one tube should be open at any given moment during pipetting steps. All other tubes and vials should be
kept closed and physically separated from the one being handled.
o
o
o
o
o
o
o
o
o
Do not pipette any of the materials by mouth.
Do not smoke, eat or drink in areas in which specimens or kit reagents are handled.
Use disposable gloves when working with clinical material possibly containing target-nucleic acids. Change
gloves after contact with potentially infectious material.
Handle all materials used (including samples, reagents, pipettes etc.) cautiously as though capable of
transmitting infectious agents.
Consult a physician immediately in the event of infectious materials being ingested or coming into contact with
open lacerations, lesions or other breaks in the skin.
Immediately clean up any spillage containing potentially infectious agents with a 0,1 % (w/v) (freshly prepared)
sodium hypochlorite solution. Dispose of the cleaning material by an accepted method.
Collect used disposable materials in a container. Close and remove container after each nucleic acid isolation
procedure.
Dispose of all materials used for nucleic acid isolation as though they contain infectious agents.
Soak tube racks used during nucleic acid isolation in a detergent (e.g. Merck Extra MAOI alkaline) for at least
one hour after each nucleic acid isolation procedure.
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o
o
Clean the centrifuge rotor used for nucleic acid isolation and the inside of the centrifuge with a detergent directly
after use, rinse with water and dry.
®
Waste from the nucleic acid release procedures using NucliSENS Lysis Buffer should be disposed of
according to local requirements.
Consult any individual instrument user manuals provided by the manufacturer for additional information regarding the
following:
o Installation and special requirements
o Operation principles, instructions, precautions, and hazards
o Manufacturer's specifications and performance capabilities
o Service and maintenance information
Materials Needed but not Provided
®
o Table-top centrifuge suitable for 2 ml NucliSENS Lysis Buffer tubes, capable of 1500 g.
o Tubes with cap for specimen storage.
o Calibrated micropipettes, volume range 10 to 1000 µl. Note: Use two different pipettes for the removal of
supernatant and the addition of wash fluid.
o Sterile, disposable aerosol resistant tips.
o Timer.
o Vortex.
o Absorbent tissue.
o Disposable gloves.
o Sodium hypochlorite solution (5 %).
o Waste container with cap.
Nucleic Acid Release
o
Consult individual instructions for use for further details when using these reagents in conjunction with other
bioMérieux products.
Reagent Preparation
o
o
o
Pre-warm a vial for approximately 30 min (at 37 °C) before starting the procedure.
Mix thoroughly, at regular intervals, during the incubation by inverting the tube. Make sure that any crystals
present have dissolved.
Cool down to room temperature.
Nucleic Acid Release Procedure
o
Consult individual instructions for use for further details when using these reagents in conjunction with other
bioMérieux products.
®
1. Spin the NucliSENS Lysis Buffer tubes at a speed and time required to spin all fluid down to the bottom of the tubes,
e.g. for 10 seconds at 1500 g.
2. Uncap the tube containing prepared specimen.
o
o
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Transfer the amount of prepared specimen to be used as the test sample into the 2 ml NucliSENS Lysis Buffer
tube. Sample volumes ranging from 10 μl to 1000 µl may be used. Note: Ensure that tubes are labeled
appropriately in order to be able to correctly identify each sample.
Record the sample volume used.
®
3. Close tubes immediately after transfer of the specimen to the NucliSENS Lysis Buffer.
o
®
Vortex the NucliSENS Lysis Buffer tubes containing specimen.
4. Repeat steps 2 and 3 for all specimens.
®
5. Incubate the NucliSENS Lysis Buffer tubes for 10 min at room temperature to release any nucleic acids present in
the specimen.
Sample storage
®
o Samples (nucleic acids in NucliSENS Lysis Buffer) can be stored at ≤ –70 °C for up to 2 months. Samples can
only be stored for 2 days at approximately 22 °C or for 4 hours at 37 °C. Storage of individual sample types may
vary.
o Consult individual instructions for use for further details when using these reagents in conjunction with other
bioMérieux products.
Notes
o
o
®
Samples in NucliSENS Lysis Buffer should be shipped on dry ice.
Handle tubes stored at ≤ –70 °C (or on dry ice) with care; tubes can become brittle at these temperatures and
may break easily.
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LIMITATIONS
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o NucliSENS Lysis Buffer 2 ml has been manufactured with the utmost care according to stringent
manufacturing procedures. However, these reagents cannot be guaranteed to be completely free of nucleic acid
contamination, and therefore it is possible that their use may interfere in amplification and detection reactions
for
certain
specific
markers.
Please contact your local representative for support.
o bioMérieux has performed a validation for the generic procedures described here on a limited number of human
specimen types. The user is responsible for validation of the lysis procedure used in their laboratory.
®
®
o NucliSENS Lysis Buffer has limited validation when used in combination with NucliSENS Magnetic Extraction
Reagents using specific human specimens; i.e. plasma, serum, cerebrospinal fluid, sputum and feces (see
references 10 - 14).
BIBLIOGRAPHY
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
Boom R, Sol CJA, van der Noordaa J, et al: Rapid and simple method for purification of nucleic acids, Journal of
Clinical Microbiology 1990; 28:495-503.
Morré, S.A., Sillekens, P. , Jacobs, M.V. (1996). RNA amplification by nucleic acid sequence-based amplification
with an internal standard enables reliable detection of Chlamidia trachomatis in cervical scrapings and urine
samples. J.Clin.Microbiol. 34,3108-3114.
van der Vliet, G.M., Cho, S.N., Kampirapap, K. (1996). Use of NASBA RNA amplification for the detection of
Mycobacterium leprae in skin biopsies from untreated and treated leprosy patients. Int.J.Lepr.Other Mycobact.Dis
64, 396-403.
Uytendaele, M., Schukkink, R., van Gemen, B., and Debevere, J. (1994). Identification of Campylobacter jejuni,
Campylobacter coli and Camplylobacter lari by the nucleic acid amplification system NASBA. J.Appl.Bacteriol. 77,
694-701.
Cassol, S., Gill, M.J., Pilon, R., et al. (1997) Quantification of human immunodeficiency virus type 1 RNA from dried
plasma spots collected on filter paper. J.Clin Microbiol. 35, 2795-2801.
Shepard, R.N.,Schock, J.Robertson, K., et al. (2000) Quantitation of human immunodeficiency virus type 1 RNA in
different biological compartments. J.Clin.Microbiol. 38, 1414-1418.
Loens, K., Ursi, D, Ieven, M., et al (2002) Detection of Mycoplasma pneumoniae in spiked clinical samples by
nucleic acid sequence-based amplification.
J. Clin Microbiol. 40, 1339-1345.
Fox, J.D., Han, S., Samuelson, A. et al (2002) Development and evaluation of nucleic acid sequence based
®
amplification (NASBA) for diagnosis of enterovirus infections using the NucliSens Basic Kit. J. Clin Virol. 24, 117130.
Wu, S.J., Lee, E., Putvatana, R. et al (2001) Detection of Dengue Viral RNA using nucleic acid sequence-based
amplification assay. J Clin Microbiol. 39, 2794-2798.
Van Deursen, P., Telles, JN., Gonzales, R., et al., (2003). Isolation of HIV-1 RNA from plasma samples using
magnetic silica particles. J. Microbiol. Methods 55:2, 531.
Van Deursen, P., Rodrigue, M, Zintilini, C., et al., (2003). Isolation of Mycobacterium nucleic acids (RNA & DNA)
from sputum and cultured strains using magnetic silica particles. J. Microbiol. Methods 55:2, 532.
Van Deursen, P., de Bie, P., Verhoeven, A., et al., (2003).Nucleic acid isolation using magnetic silica particles.
Poster 71c at the European Meeting of Molecular Diagnostics, Scheveningen, The Netherlands.
Bartels, L., Onland, G., Boel, CHE., van den Brule AJC. (2003). Isolation of Enterovirus RNA from clinical samples
using magnetic silica particles. J. Microbiol. Methods 55:2, 535.
Loens, K., Beck, T., van Deursen, P., et al., (2003). Comparison of magnetic and conventional Boom RNA
extraction using respiratory samples. J. Microbiol. Methods 55:2, 509.
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INDEX OF SYMBOLS
Symbol
Meaning
Catalog number
In Vitro Diagnostic Medical Device
Manufacturer
Temperature limit
Use by date
Batch code
Consult Instructions for Use
Contains sufficient for <n> tests
Protect from light
Date of manufacture
AVAILABILITY
Product
Catalogue number
Sufficient for
®
200293
48 samples
®
200292
48 samples
NucliSENS Magnetic Extraction Reagents
NucliSENS Lysis Buffer (2 ml)
®
The isolation, amplification and detection technologies used in the NucliSENS system are covered by various patents
and patent applications owned by or licensed to bioMérieux.
REVISION HISTORY
Change type categories :
N/A
Not applicable (First publication)
Correction
Correction of documentation anomalies
Technical change
Addition, revision and/or removal of information related to the product
Administrative
Implementation of non-technical changes noticeable to the user
Note :Minor typographical, grammar, and formatting changes are not included in the revision history.
Release date
2015/01
Part Number
14900G
Change Type
Administrative
Technical change
Change Summary
Index of symbols
Creation of the revision history table
Reagents
200292 - Package Insert – 14900G – 2015-01
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companies.
Any other name or trademark is the property of its respective owner.
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