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

Brand: Thermo Fisher Scientific

Model: PrioCHECK MAEDI-VISNA & CAEV Ab Serum Kit

Type: Operating instructions

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

For descriptions of symbols on product labels or product documents, go to thermoﬁsher.com/symbols-

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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use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses.

and its subsidiaries unless otherwise speciﬁed.

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15 December 2022

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

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

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

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