Thermo Fisher Scientific BOVIGAM Instruction Manual

Catalog Number 7600100

Pub. No. MAN0017011 Rev. A.0

NOT FOR SKIN TESTING

WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and

WARNING! POTENTIAL BIOHAZARD.

Positive

OD PC-EC

−

Nil Antigen

≥

0.1

Negative

OD PC-EC

−

Nil Antigen

0.1

OD PWM − Nil Antigen ≥ 0.5

INSTRU C TION S FOR U S E

BOVIGAM™ PC-EC Stimulating Antigen

For the laboratory diagnosis of tuberculosis in animals by the Bovine Gamma Interferon Test

gloves. Safety Data Sheets (SDSs) are available from thermofisher.com/support.

appropriate protective eyewear, clothing, and gloves.

Introduction

Applied Biosyste ms™ BOVIGAM™ PC-EC Stimulating Antigen is a stimulation antigen containing synthetic peptides. BOVIGAM™ PC-EC Stimulating Antigen is based on a peptide composition exclusively derived from ESAT-6 and C FP-10. The reconstituted solution does not contain any preservatives.

General instructions

It is essential that all blood sa mples collected for BOVIGAM™ 2G TB Kit testing are taken from a cleaned site into a tube containing heparin as an anticoagulant. Gently invert tubes several times to ensure complete mixing of blood and heparin. Blood samples should be transported to the testing laboratory at ambient temperatures (22±3°C, avoid extremes) and used in the BOVIGAM

IMPORTANT! Blood must not be stored in a refrigerator.

Test principle

The BOVIGAM™ 2G TB Kit is a rapid, blood-based assay of cell mediated immunity (CMI) for the diagnosis of bovine tuberculosis in cattle and other ruminants. BOVIGAM

™

PC-EC Stimulating Antigen is mixed with lymphocytes in

whole blood and incubated overnight. Blood plasma is then collected from above the cultures and assayed for interferon-γ (IFN-γ) production using BOVIGAM

™

2G TB Kit.

General precautions

Laboratory safety

National Safety Regulations must be strictly followed.

Test procedure

For stimulation in 96-well format (recommended)

One vial of freeze-dried BOV IGAM reconstituted with 3 mL of deionized or distilled water.

Ensure complete resolubilization. Reconstituted BOVIGAM Stimulating Anti gen may be aliquoted and stored at −80°C for several months, but must be brought to room temperature (22±3°C) and mixed thoroughly before use.

Mix blood samples evenly immediately before use.

For testing, dispense 250 µL aliquots of bloo d into culture vessels (depending on the stimulation set up you are planning: BOVIGAM Stimulating Anti gen, NIL, PWM). If more plasma is needed in later BOVIGAM

™

2G TB Kit assays, then duplicate stimula tion can be set up.

To one aliquot, add 25 µL of BOVIGAM mix thoroughly. To the other aliquots, add 25 µL of sterile PBS to one and 25 µL of PWM to the other (if a PWM control is used). Aseptic procedures should be followed using a sterile pipetting apparatus.

™

PC-EC Stimulating Antigen is

Interpretation

1. Calculate mean NIL antigen and BOVIGAM

Antigen absorbance values for each sample.

2. Compare the mean absorbance values of the NIL antigen and the

BOVIGAM

3. Blood pla sma collected from cattle, having an OD value greater than

0.100 above that of the NIL antigen, indicates the presence of Mycobacterium bovis infection.

™

PC-EC Stimulating Antigen samples for each animal.

Read the biological hazard safety information at this product’s page at thermofisher.com. Wear

™

2G TB Kit assay within 30 hours of collection.

Stimulation with PWM (stimulation control)

1. Calculate mean NIL antigen and PWM absorbance values for each

sample.

2. Compare the mean absorbance values of the NIL antigen and the PWM

samples for each animal.

Whole blood samples, which show (after PWM stimulation within the EIA analysis) a mean OD value (after subtraction of the NIL OD value) of greater than 0.5 contain vital lymphocytes.

Samples measured with lower values may indicate that the vitality of lymphocytes was decreased by transport or inappropriate handling.

Furthermore, PWM samples of stressed or immunosuppressed animals or animals in bad condition may show mean OD values below 0.5. Samples which do not reach this cut-off should not be analyzed.

CAUTION

™

PC-EC

Immunosuppression caused by recent dexamethasone treatment or parturitio n may depress IFN-γ responses to mycobacterial antigens. Animals that have received an injection of dexamethasone within one week, or that have calved within 4 weeks, should be retested to reduce the possibility of a false-negative result.

As with any biological test, this test may give a false positive or false negative result due to local conditions. A test should be interpreted in the

™

PC-EC

context of all available clinical, historical, and epidemiolog ical information relevant to the animal(s) under test. Further confirmatory testin g may be required in certain circumst ances.

™

PC-EC Stimu lating Antigen and

Responsibility for the test interpretation and consequent animal husbandry decisions rests solely on the user, and any consulting veterinarian and appropriate health advisors or authorities.

Thermo Fisher Scientific accepts no responsibility for any loss or damage, howsoever caused, arising from the interpretation of the test results.

™

PC-EC Stimulating

Retesting

Retesting may be performed at any time after the last test.

Storage

Store, protected from light, at minimum −20°C, better down to −80°C (lyophilized). Reconstitute d and aliquoted BOVIGAM Antigen should be stored frozen at −80±5°C.

Mode of issue

BOVIGA M™ PC-EC Stimulating An tigen is available in 3 mL vials.

™

PC-EC Stimu latin g

For Veterinary Use Only. For In Vitro Use Only.

Rev.

Date

Descript ion

New document. Converted the legacy document [7600100_EN.doc

trademarks, and logos.

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The information in this guide is subject to change without notice. DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S)

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Revision history of Pub. No. MAN0017011 (English)

A.0 2 October 2020

Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses.

version 1.2] to the current document template, with associated updates to the pu blication number, limited license information, warranty,

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2 October 2020

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