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cattletype BHV1 gE Ab

For the detection of antibodies to glycoprotein E of bovine herpesvirus 1 (BHV1) in samples from cattle

  • Sensitive and specific BHV1 gE-blocking ELISA
  • Suitable for serum, plasma and milk samples
  • User-friendly protocol, easy-to-use, automatable
  • Easy to integrate into existing workflows
  • Available in versatile strip or plate formats

cattletype
BHV1 gE Ab is a sensitive and specific blocking ELISA for detecting antibodies to glycoprotein E of bovine herpesvirus 1 (BHV1) in serum, plasma and milk samples from cattle. It can be used with individual samples or with pooled milk samples after sample preparation with the cattletype Milk Prep Kit.
Cat No./ID: 270203
cattletype BHV1 gE Ab (5)
For 480 reactions: 5 Test plates (strips), Wash Solution, Sample Diluent, Positive Control, Negative Control, Anti-gE-HRP Conjugate, TMB Substrate Solution, Stop Solution
Cat No./ID: 270205
cattletype BHV1 gE Ab (20)
For 1920 reactions: 20 Test plates (solid), Wash Solution, Sample Diluent, Positive Control, Negative Control, Anti-gE-HRP Conjugate, TMB Substrate Solution, Stop Solution
cattletype BHV1 gE Ab is for veterinary use only. In Germany: Licensed in accordance with §17c of the German Law on Animal Diseases (FLI-B 664).
Performance

cattletype BHV1 gE Ab enables differentiation of infected from vaccinated animals (DIVA). DIVA is possible when marker vaccines are used. The combination of cattletype BHV1 gB Ab and cattletype BHV1 gE Ab enables complete surveillance of herds (see Table 1).


Table 1. Complete surveillance of herds
 Healthy non-vaccinated animal  IBR infected cow  Traditional vaccine IBR marker vaccine
cattletype BHV1 gB Ab  Negative  Positive  Positive  Positive
cattletype BHV1 gE Ab  Negative  Positive  Positive  Negative

Further investigation on reference panels shows that cattletype BHV1 gE enables improved differentiation between infected animals and vaccinated animals than the commercial assay from Supplier I (see Table 2).

Table 2. cattletype BHV1 gE outperforms an assay from Supplier I
 
                            Sample origin
          cattletype BHV1 gE Commercial assay from Supplier I
 OD[MV]     PI
[MV]        
 Result  P/N Result
R32: Berta; negative  1.183  –22.2%  Negative  1.048  Negative
R69: TV DK, control animal, field infected  0.204  78.9%  Positive  0.125  Positive
R31: Alma, negative  1.114  –15.1%  Negative  1.054  Negative
R66: TV DK, vaccinated + infected (marker vaccinated & field infected)  0.429  55.8%  Positive  0.465  Positive
R71: TV DK, vaccinated + very early infected (marker vaccinated and field vaccinated)  0.851  –2.3%  Negative  0.931  Negative
R3: R1, 1:4, field-infected  0.413  57.3%  Positive  0.681  Suspect
R2: R1, 1:2, field-infected  0.344  64.5%  Positive  0.637  Suspect
R3: R1, 1:4, field-infected  0.406  58.1%  Positive  0.676  Suspect
R1: SH2 dilution in DCS, field infected  0.188  80.6%  Positive  0.401  Positive
R51 / 48 / 42: marker vaccinated  1.031  –6.5%  Negative  0.927  Negative
Principle
BHV1 is the causative agent of Infectious Bovine Rhinotracheitis (IBR) – a respiratory disease that causes tracheitis, rhinitis and fever. In addition, BHV1 infection can cause Infectious Pustular Vulvovaginitis (IPV), balanoposthitis and abortions. Clinical disease is often followed by latent BHV1 infection. Reactivation of the virus can cause the infection to spread in the herd.

Standard serological methods cannot distinguish between naturally infected and vaccinated animals – with the exception of IBR vaccines – which do not contain gE viral protein and therefore allow serological differentiation. cattletype BHV1 gE Ab specifically detects antibodies to gE and does not react with antibodies from gE-deleted vaccines. Therefore, this method identifies animals that have been infected with BHV1 field strains or vaccinated with non-gE-deleted IBR vaccines.

cattletype BHV1 gE Ab is a blocking ELISA. The Test Plate is coated with inactivated BHV1 antigen. During sample incubation, BHV1-specific antibodies bind to the immobilized antigen. Unbound material is removed by rinsing. The HRP-labeled, gE-specific monoclonal antibody conjugate is added, which cannot bind to the BHV1 antigen while its antigenic determinant is blocked by antibodies in the test sample. Unbound anti-gE-HRP conjugate is rinsed out. A color reaction is initiated by adding the substrate solution and stopped after 10 minutes. The optical density (OD) is measured.

 

Procedure

Serum and plasma samples can be tested after dilution. Milk samples can be tested after skimming. After sample preparation with the cattletype Milk Prep, it is also possible to test pooled milk samples.

Samples are incubated overnight with the test plate. If antibodies to glycoprotein E of BHV1 are present, they will bind to the antigen on the plates. Unbound material is removed by rinsing. 

The anti-gE-HRP conjugate is added. It contains an HRP-labeled, gE-specific monoclonal antibody. It will only bind to antigen that is not occupied by antibodies. Unbound conjugate is rinsed out.

The colour reaction is started by adding the TMB Substrate Solution and stopped after 10 minutes. The optical density (OD) is measured using a spectrophotometer. The blocking value (percentage of inhibition) is calculated from the OD values obtained with the test sample and the Negative Control, which contains no BHV1-specific antibodies. This blocking value is easy to interpret.

Applications
cattletype BHV1 gE Ab is an enzyme immunoassay (ELISA). It is intended for the detection of antibodies to glycoprotein E of bovine herpesvirus 1 (BHV1) in serum, plasma and milk samples from cattle infected with BHV1.

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