PRAXISDIENST 192679 NucleoProtect VET Stabilization and Inactivation Reagent User Manual
- June 8, 2024
- PRAXISDIENST
Table of Contents
- PRAXISDIENST 192679 NucleoProtect VET Stabilization and Inactivation
- Components
- Product description
- Product storage conditions
- Safety instructions
- Nucleic acid extraction
- Disposal
- Contact information
- Ordering information
- 9 Product use restriction / warranty
- Read User Manual Online (PDF format)
- Download This Manual (PDF format)
PRAXISDIENST 192679 NucleoProtect VET Stabilization and Inactivation
Reagent
Components
Product content
It is strongly recommended to read the protocol details in this user manual
when using NucleoProtect® VET stabilization and inactivation reagent for the
first time.
All technical literature is available online at www.mn‑net.com.
Consumables and Equipment to be supplied by the user
- Depending on the type of specimen to be collected, appropriate leak-proof screw cap tubes, swabs, blood collection tubes or cannulas.
- Personal protection equipment (e.g., lab coat, gloves, goggles).
Product description
The basic principle
The NucleoProtect® VET stabilization and inactivation reagent is intended for
the collection of blood or swab specimens from animals, inactivate viral
pathogens and preserve viral nucleic acids (DNA and RNA) by inactivating
nucleases. The NucleoProtect® VET stabilization and inactivation reagent
allows the preservation and save transport of veterinary whole blood or swab
samples from the site of sample collection site to the analyzing laboratory.
The NucleoProtect® VET stabilization and inactivation reagent allows the safe
collection and transport of whole blood or swab specimens from animals and
subsequent viral nucleic acid (RNA/DNA) extraction for use in downstream
analyses such as (RT)-PCR or q(RT)-PCR.
Product specification
The NucleoProtect® VET reagent was tested with animal blood collected with
NucleoProtect® VET Blood Tubes (REF 740755) and EDTA-anticoagulated animal
blood. The user is responsible to validate other anticoagulants used in
conjunction with the NucleoProtect® VET reagent. The reagent was also tested
with different swab materials including foam tip swabs, cotton fibers,
synthetic fibers and flocked swab. The user is responsible to validate the
individual swab used in conjunction with the NucleoProtect® VET reagent. The
product is not intended for human use. The product is not intended for human
use.
Inactivation of viruses
The inactivating of high risk viruses was tested in different background
matrices (see table below) using the Tissue Culture Infectious Dose (TCID)
assay. Viruses were spiked into the different matrices with titers between 10⁶
and 10⁷ virus particles per mL and incubated for 30 minutes in the
NucleoProtect® VET reagent. Samples were diluted to prevent toxic effects of
the NucleoProtect® VET reagent on cell culture cells and virus specific cell
culture cells were infected with the treated viruses. Culture cells were
subsequently incubated several days and analyzed for cytopatic effects (CPE).
An aliquot of each cell culture supernatant was then passaged to a fresh cell
culture. In case of a lack of viral growth in both cell culture infection
assays the viruses were considered as completely inactivated. The titer
reduction was calculated from the starting titer and the dilution factor.
The level of virus titer reduction is shown in the table below (data kindly
provided by the Friedrich-Loeffler-Institut, Federal Research Institute for
Animal Health of Germany).
Virus
| Background Matrix|
Reduction of virus titer
|
---|---|---|---
Foot-and-mouth disease virus (FMDV)|
Bovine tongue ephithelium
| |
Bluetongue virus (BTV-5)|
Whole blood Raw milk
Saliva
| > 10⁴ (corresponds to an inactivation capacity of
99,99 %)
|
Peste des petits ruminants virus (PPRV)|
Lumpy skin disease virus (LSDV)| | |
Bovine Coronavirus (BCoV)| Saliva| |
The inactivation capacity of the NucleoProtect® VET reagent has been
demonstrated for various background matrices and viruses known to have
different envelope structures as well as susceptibilities to disinfectants:
FMDV (small non-enveloped virus), BTV-5 (large non-enveloped virus), LSDV
(large enveloped virus), PPRV and BCoV (enveloped virus).
Claims for inactivation of other viruses or background matrices cannot be
made. Samples must be incubated for at least 30 min for complete inactivation.
Sample stability
The stabilization capability of the NucleoProtect® VET reagent was tested with
all naturally occurring viral genome structures: ss (+ / -)RNA, dsRNA and
dsDNA. In particular, the following viruses were tested:
Virus Genome structure
Foot-and-mouth disease virus (FMDV) Porcine enterovirus (PEV)
Bovine viral diarrhea virus (BVDV)
| ss(+)RNA
Peste des petits ruminants virus (PPRV) Canine distemper virus (CDV)| ss(-)RNA
Bluetongue virus (BTV serotypes 5 and 8)| dsRNA
Lumpy skin disease virus (LSDV) Vaccinia virus (VV)
Bovine herpes Virus (BoHV)
| dsDNA
Stabilization of viral RNA and DNA was tested with spiked blood, or swab samples using virus specific q(RT)-PCR assays. NucleoProtect® VET-stabilized samples (swabs / blood) can be stored and shipped at the following conditions without sensitivity loss*:
Storage condition Storage time*
15 – 25 °C| 7 days
4 °C| 7 days
37 °C| 2 days
-20 °C| 30 days
- the specified storage time indicates how long the samples can be stored at the specified condition with a change in Cq-value of < 1 on average, as demonstrated in q(RT)-PCR analyses comparing stored samples to the time point of collection.
The NucleoProtect® VET reagent is not intended for preserving morphology or viability of microorganisms. Due to its lytic and denaturing activity, the NucleoProtect® VET reagent is not suited for serological testing.
Handling of sample material
The NucleoProtect® VET reagent is intended for safe collection and transport
of animal whole blood or swab samples only.
The NucleoProtect® VET stabilization reagent appears colorless and clear. Do
not use NucleoProtect® VET reagent if the solution appears discolored or
contains precipitates.
Do not use if the product or packaging is damaged or if there is evidence of
leakage.
Recommended NucleoProtect® VET reagent volume for different sample types.
Sample Recommended Volume
Animal whole blood| Add 2.5 volumes of NucleoProtect® VET reagent per volume of animal whole blood.
Calculate the required NucleoProtect® VET reagent volume accordingly for individual blood volume used, e.g.:
For 100 µL blood add 250 µL NucleoProtect® VET reagent For 500 µL blood add 1250 µL NucleoProtect® VET reagent For 1.0 mL blood add 2.5 mL NucleoProtect® VET reagent For 1.5 mL blood add 3.75 mL NucleoProtect® VET reagent
Swabs| The amount of NucleoProtect® VET reagent needed for effective sample preservation and inactivation depends on the size and material of the swab (swab head material, such as cotton, or synthetic / inert fibres).
We recommend adding enough NucleoProtect® VET reagent to submerge the swab material. Ensure that the swab material is compatible with the NucleoProtect® VET reagent.
The user is responsible to use leak proof container in conjunction with the
NucleoProtect® VET reagent.
The NucleoProtect® VET Blood Tube is intended for professional use only.
Follow veterinarian and laboratory’s standard procedures for collecting blood
or swab samples from animals and apply appropriate safety procedures.
Collecting Swab samples
Insert Swab into the NucleoProtect® VET reagent after collecting the
veterinary sample. Do not insert the swab into the NucleoProtect® VET reagent
before collecting the sample!
Collecting Blood samples
Follow veterinary and laboratory standard procedures for collecting blood from
animals and apply appropriate safety practices when collecting blood. If other
primary blood collection tubes than EDTA-tubes are used to collect blood from
animals, the user is responsible to validate the use in conjunction with the
NucleoProtect® VET reagent.
Product storage conditions
Store NucleoProtect® VET reagent at ambient temperatures (15 – 25 °C).
The product can be used until the expiry date printed on the label.
Safety instructions
Always wear suitable protective clothing (e.g., protective clothes, gloves,
and goggles) when working with the NucleoProtect® VET reagent or when working
with potentially infectious biological material.
For more information consult the appropriate Material Safety Data Sheets (MSDS
available online at www.mn-net.com/msds).
Avoid mixing the NucleoProtect® VET reagent with sodium hypochlorite (bleach)
or other strong acids and bases. The mixtures could form highly reactive
compound when combined with bleach!
The NucleoProtect® VET reagent contains chemical additives. Prevent contact
with the animal.
Remove spilled NucleoProtect® VET reagent with an excess of water.
Nucleic acid extraction
The NucleoProtect® VET reagent is compatible with the MACHEREY-NAGEL
NucleoSpin® VET kit (silica membrane single spin kit; REF 740842.10 / .50 /
.250) and the NucleoMag® VET kit (magnetic bead based kit; REF 744200.1 / .4)
nucleic acid extraction systems.
The NucleoProtect® VET reagent is also compatible with other common commercial
nucleic acid extraction solutions. The user is responsible for validating
additional nucleic acid extraction and purification methodologies.
*Nucleic acid extraction using the NucleoMag® VET kit (see ordering
information below):**
For detailed description of the nucleic acid extraction procedure from
NucleoProtect® VET-stabilized samples using the NucleoMag® VET kit, please
refer to the user manual.
Download user manual NucleoMag® VE
www.mn-net.com/NucleoMagVETmanual
Carefully read the user manual of the NucleoMag® VET kit before starting the
nucleic acid extraction procedure.
For the processing of blood collected with the NucleoProtect® VET reagent
using the NucleoMag® VET kit, additional buffer VIA is needed (REF 744206, see
ordering information).
Non-stabilized and NucleoProtect® VET-stabilized samples can be processed in
parallel. For detailed description, please refer to the user manual.
*The NucleoMag® VET extraction kit has been automated on a wide variety of open liquid-handling or magnetic-rod platforms. Please contact MACHEREY-NAGEL for technical automation support and verified methodologies: tech-bio@mn-net.com I+49 24 21 969-270
Nucleic acid extraction using the NucleoSpin® VET kit (see ordering
information below):
For detailed description of the nucleic acid extraction procedure from
NucleoProtect® VET-stabilized samples using the NucleoSpin® VET kit, please
refer to the user manual.
Download user manuel NucleoSpin® VET:
www.mn-net.com/NucleoSpinVETmanual
Carefully read the user manual of the NucleoSpin® VET kit before starting the
nucleic acid extraction procedure.
For the processing of NucleoProtect® VET-stabilized samples using the
NucleoSpin® VET kit, additional buffer PFN is needed (REF 740121.5, see
ordering information).
Non-stabilized and NucleoProtect® VET-stabilized samples can be processed in
parallel. For detailed description, please refer to the user manual.
Disposal
Always dispose hazardous, infectious or biologically contaminated materials in
a safe and acceptable manner and in accordance with all local and regulatory
requirements.
Discard blood collection sharps like cannulas in biohazard containers approved
for their disposal. Please follow local waste regulations.
Contact information
For more detailed product use restriction / warranty please visit: www.mn-
net.com
Please contact our technical support for any product related inquiries:
MACHEREY-NAGEL Germany Tel.:
+49 (0) 24 21 969-270
e-mail: tech-bio@mn-net.com
Ordering information
| Product | Description | Pack of | REF |
|---|---|---|---|
| NucleoProtect® VET stabilization and inactivation reagent | NucleoProtect® VET |
stabilization and inactivation reagent for filling into own transport or storage vessels| 50 /
500 mL
| 740750.50 /
500
Accessory / related products
NucleoProtect® VET Blood Collection Tubes| Blood collection tube prefilled
with 4.0 mL NucleoProtect® VET stabilization and inactivation reagent,
compatible with standard Luer cannulas, box of 50 tubes| 50| 740755
---|---|---|---
NucleoProtect® VET Swab Tube| Screw cap swab collection tube prefilled with
1.5 mL
NucleoProtect® VET stabilization and inactivation reagent including swabs
| 50| 740760
Secondary caps for NucleoProtect® VET Blood Tubes Tubes| Sealing caps for
NucleoProtect® VET Blood Tubes| 100| 740756
Secondary caps for NucleoProtect® VET Swab Tubes| Sealing caps for
NucleoProtect® VET Swab Tubes| 100| 740761
NucleoMag® VET| Magnetic bead based kit for extraction of RNA and DNA from
veterinary samples, containing NucleoMag® B-Beads, buffers, Carrier RNA,
Proteinase K| 1 × 96 /
4 × 96
| 744200.1 / .4
Buffer VIA| Needed for the nucleic acid extraction from NucleoProtect® VET
collected blood samples using the NucleoMag® VET kit| 150 mL| 744206
NucleoSpin® VET| Single spin kit for extraction of RNA and DNA from veterinary
samples, containing binding columns, collection Tubes (2 mL), collection tubes
for lysis and elution (1.5 mL), buffers, liquid Proteinase K, Carrier RNA
(lyophilized)| 10 / 50 /
250
| 740842.10 /
.50 / .250
Buffer PFN| Needed for the nucleic acid extraction from NucleoProtect® VET collected samples using the NucleoSpin® VET kit| 5 mL| 740121.5
Visit www.mn-net.com for more detailed product information or contact our technical support (tech-bio@mn-net.com)
9 Product use restriction / warranty
All MACHEREY-NAGEL products are designed for their intended use only. They are not intended to be used for any other purpose. The description of the intended use of the products can be found in the original MACHEREY-NAGEL product leaflets. Before using our products, please observe the instructions for use and the safety instructions from the respective Material Safety Data Sheet of the product.
This MACHEREY-NAGEL product is carrying documentation stating specifications
and other technical information. MACHEREY-NAGEL warrants to meet the stated
specifications. The provided warranty is limited to the data specifications
and descriptions as given in the original MACHEREY-NAGEL literature. No other
statements or representations, written or oral, by MACHEREY-NAGELS employees,
agents or representatives, except written statements signed by a duly
authorized officer of MACHEREY-NAGEL are authorized. They should not be relied
upon by the costumer and are not a part of a contract of sale or of this
warranty.
Liability for all possible damages that occur in any connection with our
products is limited to the utmost minimum as stated in the general business
terms and conditions of MACHEREY-NAGEL in their latest edition which can be
taken from the company’s website. MACHEREY-NAGEL does not assume any further
warranty.
Products and their application are subject to change. Therefore, please
contact our Technical Service Team for the latest information on MACHEREY-
NAGEL products. You may also contact your local distributor for general
scientific information. Descriptions in MACHEREY-NAGEL literature are provided
for informational purposes only.
Contact MN
Germany and international
MACHEREY-NAGEL GmbH & Co. KG
Valencienner Str. 11 · 52355 Düren · Germany
Tel. : +49 24 21 969-0
Toll-free: 0800 26 16 000 (Germany only)
E-mail: info@mn-net.com
Technical Support Bioanalysis
Tel.: +49 24 21 969-270
E-mail: tech-bio@mn-net.com
USA
MACHEREY-NAGEL Inc.
924 Marcon Blvd. · Suite 102 · Allentown PA, 18109 · USA
Toll-free: 888 321 6224 (MACH)
E-mail: sales-us@mn-net.com
France
MACHEREY-NAGEL SAS
1, rue Gutenberg – BP135 · 67720 Hoerdt Cedex · France
Tel.: +33 388 68 22 68
E-mail: sales-fr@mn-net.com
MACHEREY-NAGEL SAS (Société par Actions Simplifi ée) au capital de 186600 €
Siret 379 859 531 00020 · RCS Strasbourg B379859531 · N° intracommunautaire
FR04 379 859 531
Switzerland
MACHEREY-NAGEL AG
Hirsackerstr. 7 · 4702 Oensingen · Switzerland
Tel.: +41 62 388 55 00
E-mail: sales-ch@mn-net.com
www.mn-net.com
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