Thermo Fisher Scientific BOVIGAM PC-HP Stimulating Antigen Operating instructions

Brand: Thermo Fisher Scientific

Model: BOVIGAM PC-HP Stimulating Antigen

Type: Operating instructions

For Veterinary Use Only. For In Vitro Use Only.

INSTRUCTIONS FOR USE

BOVIGAM™ PC-HP Stimulating Antigen

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

Catalog Number 7600105

Pub. No. MAN0017013 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

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

WARNING! POTENTIAL BIOHAZARD.

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

appropriate protective eyewear, clothing, and gloves.

Introduction

Applied Biosystems™ BOVIGAM™ PC-HP Stimulating Antigen is a stimulation antigen containing synthetic peptides. BOVIGAM™ PC-HP Stimulating Antigen

is based on a peptide composition exclusively derived from ESAT-6 and CFP-10, RV3615c, OmpA, and two additional antigens. The reconstituted solution does

not contain any preservatives.

General instructions

It is essential that all blood samples 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™ 2G TB Kit assay within 30 hours of collection.

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-HP 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

the 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 BOVIGAM™ PC-HP Stimulating Antigen is

reconstituted with 3 mL of deionized or distilled water.

Ensure complete resolubilization. Reconstituted BOVIGAM™ PC-HP

Stimulating Antigen 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 blood into culture vessels

(depending on the stimulation set up you are planning: BOVIGAM™ PC-HP

Stimulating Antigen, NIL, PWM). If more plasma is needed in later

BOVIGAM™ 2G TB Kit assays, then duplicate stimulation can be set up.

To one aliquot, add 25 µL of BOVIGAM™ PC-HP Stimulating Antigen and

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.

Interpretation

1. Calculate mean NIL antigen and BOVIGAM™ PC-HP Stimulating

Antigen absorbance values for each sample.

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

BOVIGAM™ PC-HP Stimulating Antigen samples for each animal.

Positive

OD PC-HP

−

Nil Antigen

≥

0.1

Negative

OD PC-HP

−

Nil Antigen

0.1

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

0.100 above that of the NIL antigen, indicates the presence of

Mycobacterium bovis infection.

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.

OD PWM − Nil Antigen ≥ 0.5

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

Immunosuppression caused by recent dexamethasone treatment or

parturition 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

context of all available clinical, historical, and epidemiological information

relevant to the animal(s) under test. Further confirmatory testing may be

required in certain circumstances.

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.

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). Reconstituted and aliquoted BOVIGAM™ PC-HP Stimulating

Antigen should be stored frozen at −80±5°C.

Mode of issue

BOVIGAM™ PC-HP Stimulating Antigen is available in 3 mL vials.

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2 October 2020

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The information in this guide is subject to change without notice.

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Revision history of Pub. No. MAN0017013 (English)

Rev.

Date

Description

A.0 2 October 2020

New document. Converted the legacy document [7600105_EN.doc

version 1.2] to the current document template, with associated updates

to the publication number, limited license information, warranty,

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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.

Fisher Scientific and its subsidiaries unless otherwise specified.

Thermo Fisher Scientific BOVIGAM PC-HP Stimulating Antigen Operating instructions

Thermo Fisher Scientific BOVIGAM PC-HP Stimulating Antigen Operating instructions

Thermo Fisher Scientific BOVIGAM PC-HP Stimulating Antigen Operating instructions

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