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Thermo Fisher Scientific PrioCHECK MAEDI-VISNA & CAEV Ab Serum Kit Operating instructions

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PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit
ELISA for in vitro detection of antibodies against Maedi-Visna virus and Caprine
Arthritis Encephalitis virus in serum of ruminants
Catalog NumberVETCAEV5
Pub.No. MAN0028311 Rev. A.0
IMPORTANT! This product is registered with the French Reference Laboratory – Anses
Maisons-Alfort. Specific requirements apply for the use of the kit for diagnostic procedures
in France, see Appendix A, “Recommended protocol in France”.
Technique Species Sample matrices Test type
Single–well blocking ELISA in strip plates
Caprine
Serum
Individual
Ovine Individual
Pooled samples (France only)
WARNING! Read the Safety Data Sheets (SDSs) and follow the handling
instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety Data
Sheets (SDSs) are available from thermofisher.com/support.
WARNING! BIOHAZARD. Read the biological hazard safety information at this
product’s page at thermofisher.com. Follow all applicable local, state/provincial,
and/or national regulations for working with biological samples.
Product description
The Applied Biosystems™ PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit
(Cat.No.VETCAEV5) is an ELISA that enables detection of anti-MVV/CAEV antibodies of
Maedi-Visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) in ruminant serum.
Caprine arthritis encephalitis virus (CAEV) is a disease of caprines caused by a lentivirus. The
associated clinical signs include:
• Arthritis and mastitis in adults
• Encephalitis in young animals
Maedi-Visna virus (MVV) is a disease of ovines also caused by a lentivirus. The associated
clinical signs include:
• Respiratory diculties or dyspnea
• Arthritis
Serological diagnosis for these two diseases must be performed to certify a caprine and ovine
herd. Clinical signs are only expressed in some seropositive animals.
INSTRUCTIONS FOR USE
For Veterinary Use Only. For In Vitro Use Only.
Test principle
The kit is based on the principle of a blocking ELISA.
1. Samples and controls are added to the MVV/CAEV antigen-coated microtiter plate. If
they are present, anti-MVV/CAEV-specific antibodies bind to the antigen and form antigen-
antibody complexes.
2. After washing, a monoclonal anti-gp135 HRP-labeled conjugate is added, which binds to
any antigen that remains free on the plate.
3. Unbound conjugate is removed by washing before the addition of a chromogen substrate.
A blue color results from substrate oxydation by the HRP-conjugate.
4. After stopping the reaction, the color remains blue. Results are read by an ELISA plate
reader.
Positive samples will show reduced or no staining.
Contents and storage
Catalog numbers that appear as links open the web pages for those products.
This kit contains sucient components for 480 tests.
Table1PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit (Cat.No.VETCAEV5)
Component Amount Storage
1–Coated microplate MVV/CAEV[1] 5 units 2–8°C[2]
2–Negative Control MVV/CAEV (NC) 1.8mL 2–8°C
3–Positive Control MVV/CAEV (PC) 1.8mL
4–Conjugate (100x) MVV/CAEV anti-gp135 HRP 100x concentrate 500µL 2–8°C[3]
A–Wash (10x) MVV/CAEV concentrate 240mL
2–8°C
B1–Sample DB MVV/CAEV (green) 20mL
B2–Conjugate diluent MVV/CAEV 50mL
C–Substrate solution MVV/CAEV 60mL
D–Stop solution MVV/CAEV 60mL
Adhesive plate films 10 RT[4]
[1] MVV/CAEV antigen-coated plate, 12 strips of 8 wells
[2] Seal any unused strips in a plastic bag with the provided desiccant sachet, then store at 2–8°C until the expiration
date of the kit.
[3] The diluted conjugate solution must be used immediately after preparation.
[4] Room temperature is defined as 17–25°C.
2 PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use
Required materials not supplied
Unless otherwise indicated, all materials are available through thermofisher.com. "MLS"
indicates that the material is available from fisherscientific.com or another major laboratory
supplier.
Item Source
Precision or multi-channel pipettes MLS
Disposable pipette tips MLS
Distilled or deionized water MLS
ELISA reader with 620-, 630-, or 650-nm filter MLS
Disposable containers MLS
Procedural guidelines
• Do not mix components from dierent kit batches.
• Use disposable supplies to prevent contamination of the reagents.
• Do not pipette by mouth.
• Store the kit and all reagents according to condition indicated on the product.
• Unused strips should be stored within a sealed plastic bag with the provided dessicant
sachet at 2–8°C.
• Wear appropriate personal protection equipment: gloves, cover, glasses.
• Properly decontaminate and eliminate all wastes in accordance with local regulations.
• Throughout this user guide room temperature is defined as 17–25°C.
• The tolerance for incubation times is ±10%.
Before you begin
Prepare sample
Fresh, refrigerated (8 days at 2–8°C), or frozen (1 year at <−16°C) serum may be used.
Individual sera are tested without dilution. The test requires 50 μL per well of each sample or
control.
PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use 3
Prepare the reagents
• The following reagents are ready for use: 1‑Coated microplate MVV/CAEV, 2‑Negative
Control MVV/CAEV, 3‑Positive Control MVV/CAEV , B1‑Sample DB MVV/CAEV,
B2‑Conjugate diluent MVV/CAEV, C‑Substrate solution MVV/CAEV, and D‑Stop
solution MVV/CAEV.
• A‑Wash (10x) MVV/CAEV must be diluted to 1:10 with distilled or deionized water. The
diluted wash solution can be kept for 8 days at 2–8°C.
Example: For one strip, add 3mL of the A‑Wash (10x) MVV/CAEV to 27 mL of water.
For one plate, add 35mL of the A - Wash (10x) MVV/CAEV to 315 mL of water. Mix after
dilution.
Note: Due to the high concentration of salts, A‑Wash (10x) MVV/CAEV can contain
crystals. Before dilution, shake the bottle to dissolve any crystals.
• 4‑Conjugate (100x) MVV/CAEV must be diluted to 1:100 with the B2‑Conjugate diluent
MVV/CAEV. Mix after dilution. Use the diluted Conjugate MVV/CAEV immediately after
preparation.
Perform the ELISA procedure
IMPORTANT! Specific requirements apply for the use of the kit in diagnostic procedures in
France, see Appendix A, “Recommended protocol in France”.
Note: All reagents should be equilibrated to room temperature before use. It is recommended
to use disposable containers for distribution (diluted Wash, diluted Conjugate MVV/CAEV,
C‑Substrate solution MVV/CAEV, and D‑Stop solution MVV/CAEV).
Use undiluted samples and controls.
Note: After opening a new plastic bag, it is
recommended to immediately put the unused plate or
strips in a plastic bag with the provided desiccant
sachet. Store protected from light.
1. Add 50μL of the 3‑Positive Control MVV/CAEV
reagent to the appropriate wells of the assay plate
(for example, add to A1 and B1 wells).
2. Add 50μL of the 2‑Negative Control MVV/CAEV
reagent to the appropriate wells of the assay plate
(for example, add to C1 and D1 wells).
3. Add 50μL of the test samples to the remaining
wells.
4. Tap or gently agitate each of the long sides of the
loaded assay plate to make sure the samples coat
the bottom of the wells.
5. Cover the plate with an adhesive plate film.
1Incubate the samples
and controls
4 PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use
1Incubate the
samples and
controls (continued)
6. Incubate the plate at room temperature depending
on your application.
• 1 hour, for a CAEV application
• 2 hours, for a MAEDI-VISNA application
For simultaneous analysis of CAEV and MAEDI-
VISNA: use the same plate by adding the MAEDI-
VISNA controls and samples first. After 1 hour, add
the CAEV controls and samples. This application
can be used on a ELISA automated analysis (type
BEP, TECAN).
7. Empty the plate, then wash 5 times with 300μL of
the diluted wash solution per well.
This washing step may be manual or automated
using a plate washer.
8. Empty the plate, then tap the plate on a paper towel
to remove any traces of liquid. Do not let the plate
dry.
1. Add 50μL of the diluted Conjugate MVV/CAEV to
each well.
2. Mix gently, then cover the plate with a new
adhesive plate film.
3. Incubate the plate at room temperature for
30 minutes.
4. Empty the plate, then wash 5 times with 300μL of
the diluted wash solution per well.
This washing step may be manual or automated
using a plate washer.
5. Empty the plate, then tap the plate on a paper towel
to remove any traces of liquid. Do not let the plate
dry.
2Add the diluted
Conjugate MVV/CAEV
1. Add 50μL of the C‑Substrate solution MVV/CAEV
to each well. Mix the plate gently for 2seconds.
2. Incubate the plate at room temperature for
20 minutes protected from light. Do not cover the
plate.
3. Add 50 μl of the D‑Stop solution MVV/CAEV to
each well in the same order as the C‑Substrate
solution MVV/CAEV. Mix the plate gently for
2seconds.
4. Wipe the bottom surface of the plate with a soft
tissue.
5. Read the plate at a wavelength of 620‑nm, 630‑nm,
or 650‑nm on a microplate reader within 30 minutes
of stopping the reaction.
3Perform the substrate
reaction, then read the
plate
PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use 5
Analize the results
Calculate the results
Calculate the mean OD of the PC (ODm PC), and of the NC (ODm NC).
For each sample, calculate the percentage inhibition (% Inh):
% Inh = [ (ODm NC − OD Sample) / ODm NC ] × 100
Note: For the negative samples, it is possible to obtain % Inh < 0.
Validation criteria
The test is validated if:
• ODm NC > 0.300
• % Inh PC ≥ 35 %
If the above criteria are not met, read the instructions again, then repeat the test. If the results
are still invalid, please contact technical support.
Results interpretation
For the diagnosis of CAEV and MAEDI-VISNA:
Results Interpretation
% Inh <35 Negative
% Inh ≥35 Positive
AppendixARecommended protocol in France
This product is registered with the French Reference Laboratory - Anses Maisons-Alfort, for its
use in France. This protocol complies with French Administrator requirements.
Before you begin
Prepare samples and controls
Fresh, refrigerated (8 days at 2–8°C), or frozen (1 year at <−16°C) serum may be used.
The pre-homogenized individual samples and controls are tested at a 1:2 dilution with the
B1–Sample DB MVV/CAEV.
Tests of pools of up to 5 sera are possible. Prepare the pooled samples as follows.
1. Pool sera at equal volumes.
2. Dilute the sample by adding 25μL of the B1–Sample DB MVV/CAEV to 25μL of the
pooled sera.
6 PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use
Perform the ELISA procedure
IMPORTANT! This product is registered with the French Reference Laboratory - Anses
Maisons-Alfort, for its use in France. This protocol complies with French Administrator
requirements.
Note: All reagents should be equilibrated to room temperature before use. It is recommended
to use disposable containers for distribution (diluted Wash, diluted Conjugate MVV/CAEV,
C‑Substrate solution MVV/CAEV, and D‑Stop solution MVV/CAEV).
Samples and controls must be diluted 1:2 with the
B1‑Sample DB MVV/CAEV as described in “Prepare
samples and controls” on page6.
Note: After opening a new plastic bag, it is
recommended to immediately put the unused plate or
strips in a plastic bag with the provided desiccant
sachet. Store protected from light.
1. Add 50μL of the diluted 3‑Positive Control
MVV/CAEV reagent to the appropriate wells of the
assay plate (for example, add to A1 and B1 wells).
2. Add 50μL of the diluted 2‑Negative Control
MVV/CAEV reagent to the appropriate wells of the
assay plate (for example, add to C1 and D1 wells).
3. Add 50μL of the diluted test samples (individual or
pooled sera) to the remaining wells.
4. Tap or gently agitate each of the long sides of the
loaded assay plate to make sure the samples coat
the bottom of the wells.
5. Cover the plate with an adhesive plate film.
6. Incubate the plate at room temperature for 1 hour.
7. Empty the plate, then wash 5 times with 300μL of
the diluted wash solution per well.
This washing step may be manual or automated
using a plate washer.
8. Empty the plate, then tap the plate on a paper towel
to remove any traces of liquid. Do not let the plate
dry.
1Incubate the samples
and controls
PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use 7
1. Add 50μL of the diluted Conjugate MVV/CAEV to
each well.
2. Mix gently, then cover the plate with a new
adhesive plate film.
3. Incubate the plate at room temperature for
30 minutes.
4. Empty the plate, then wash 5 times with 300μL of
the diluted wash solution per well.
This washing step may be manual or automated
using a plate washer.
5. Empty the plate, then tap the plate on a paper towel
to remove any traces of liquid. Do not let the plate
dry.
2Add the diluted
Conjugate MVV/CAEV
1. Add 50μL of the C‑Substrate solution MVV/CAEV
to each well. Mix the plate gently for 2seconds.
2. Incubate the plate at room temperature for
20 minutes protected from light. Do not cover the
plate.
3. Add 50 μl of the D‑Stop solution MVV/CAEV to
each well in the same order as the C‑Substrate
solution MVV/CAEV. Mix the plate gently for
2seconds.
4. Wipe the bottom surface of the plate with a soft
tissue to remove any dust.
5. Read the plate at a wavelength of 620‑nm, 630‑nm,
or 650‑nm on a microplate reader within 30 minutes
of stopping the reaction.
3Perform the substrate
reaction, then read the
plate
Analyze the results
Calculate the results
1. Calculate the mean OD of the PC (ODm PC), and of the NC (ODm NC).
For each caprine sample, calculate the percentage inhibition (% Inh):
% Inh = [ (ODm NC − ODsample) / ODm NC ] × 100
Note: For the negative samples, it is possible to obtain % Inh < 0.
2. For ovine analysis (MAEDI-VISNA), laboratories in France must follow this specific
calculation, which complies with French Administrator requirements:
% Inh = [ ( (ODm NC − ODm PC) − ODsample ) / (ODm NC − ODm PC) ] × 100
Validation criteria
The test is validated if: % Inh PC ≥ 60% and ODm NC > 0.300
8 PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use
Results interpretation
Interpret the results according to the following table.
Table2Individual serum (caprine and ovine)
Results Interpretation
% Inh <60 Negative
% Inh ≥60 Positive
Table3Pooled sera (ovine only)
Results Interpretation
% Inh <35 Negative
% Inh ≥35 Positive
AppendixBDocumentation and support
Customer and technical support
Visit thermofisher.com/support for the latest service and support information.
• Worldwide contact telephone numbers
• Product support information
– Product FAQs
– Software, patches, and updates
– Training for many applications and instruments
• Order and web support
• Product documentation
– User guides, manuals, and protocols
–Certificates of Analysis
– Safety Data Sheets (SDSs; also known as MSDSs)
Note: For SDSs for reagents and chemicals from other manufacturers, contact the
manufacturer.
Limited product warranty
Life Technologies Corporation and/or its aliate(s) warrant their products as set forth in the
Life Technologies' General Terms and Conditions of Sale at www.thermofisher.com/us/en/
home/global/terms-and-conditions.html. If you have any questions, please contact Life
Technologies at www.thermofisher.com/support.
PrioCHECK™ MAEDI-VISNA & CAEV Ab Serum Kit Instructions for Use 9
Prionics Lelystad B.V. | Platinastraat 33 | 8211 AR Lelystad | The Netherlands
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definition.
Revision history:Pub.No.MAN0028311A.0
Revision Date Description
A.0 15 December 2022 New document created for PrioCHECK™ MAEDI-VISNA &
CAEV Ab Serum Kit.
The information in this guide is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S)
WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL
DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
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