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cattletype BHV1 gB Ab

For detection of antibodies to bovine herpesvirus 1 in samples from cattle

  • Highly sensitive and specific BHV1 gB-blocking ELISA
  • Suitable for serum, plasma, and milk samples
  • User-friendly test protocol with ready-to-use colored reagents
  • Results in less than 3.5 hours
  • Available in versatile strip or plate formats
Effective April 1, 2018, product available exclusively from INDICAL BIOSCIENCE (formerly QIAGEN Leipzig). Learn more.


cattletype BHV1 gB Ab is a sensitive and specific blocking ELISA for detecting antibodies to glycoprotein B 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.

cattletype BHV1 gB Ab is for veterinary use only.

In Germany: Registered in accordance with §17c of the German Law on Animal Diseases. Registration No.: FLI-B 491

In Switzerland: Approved by the Federal Veterinary Office (BVET)
Cat No./ID: 270043
cattletype BHV1 gB Ab (5)
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For 480 reactions: 5 Test plates (strips), Wash Solution, Sample Diluent, Positive Control, Negative Control, Anti-gB-HRP Conjugate, TMB Substrate Solution, Stop Solution
Effective April 1, 2018, product available exclusively from INDICAL
Cat No./ID: 270045
cattletype BHV1 gB Ab (20)
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For 1920 reactions: 20 Test plates (solid), Wash Solution, Sample Diluent, Positive Control, Negative Control, Anti-gB-HRP Conjugate, TMB Substrate Solution, Stop Solution
Effective April 1, 2018, product available exclusively from INDICAL
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Specificity of cattletype BHV1 gB Ab.

A sample set of 579 serum and 514 milk samples from BHV1-free cattle was tested using cattletype BHV1 gB Ab. The sample incubation time was 2 hours. An inhibition value (percentage of inhibition, PI) below 45% indicates the sample is negative for antibodies to glycoprotein B of BHV1. An inhibition value above 55% indicates the sample is positive for the BHV1-specific antibodies. Only one serum sample tested false positive, providing a diagnostic specificity for this sample set of 99.9%.

Performance

cattletype BHV1 gB Ab is specific for antibodies to glycoprotein B of BHV1. A negative population of 579 serum samples and 514 milk samples from BHV1-free herds were tested. All of the samples except one tested negative for the BHV1-specific antibodies, providing a diagnostic specificity of 99.9% (see the figure Specificity of cattletype BHV1 gB Ab).

cattletype BHV1 gB Ab shows excellent sensitivity. Tests were performed using the reference sample sets from the European Ring Trial as described in Kramps et al. (2004). The serum sets contained the three European BHV1 reference serum samples (positive, weak positive, and negative), and serum samples from infected, vaccinated, vaccinated and challenged, and non-infected cattle. The test correctly detected antibodies against the virus in both the positive and weak positive reference samples and scored 100% correct across the whole reference panel (see the table Sensitivity of cattletype BHV1 gB Ab).

Sensitivity of cattletype BHV1 gB Ab
Sample designation Reference status  PI cattletype BHV1 gB Ab result
 EU1 Positive  90.7%  Positive
 EU2 Weak positive  75.6%  Positive
 EU3 Negative  0.3%  Negative

Sample set A as described in Kramps et al. (2004) was tested with cattletype BHV1 gB Ab. The incubation time was 2 hours. The percentage of inhibition (PI) is indicated.

Set Reference status  n cattletype BHV1 gB Ab result  Correct PI 
      Positive   Negative     
 B Infected  12  12    100% 89.9% 
 C Vaccinated  9  9    100% 97.5%
 D Vaccinated and challenged  6  6    100% 92.1% 
 G Non-infected  7  –   7  100% 8.7%
  BHV5-infected  1  1    100% 85.8% 

The indicated numbers of samples from sample sets B, C, D, and G as described in Kramps et al. (2004) were tested with cattletype BHV1 gB Ab. A sample from an animal infected with BHV5 was also tested. The incubation time was 2 hours. The test scored 100% correct across the whole reference panel.
Principle

Bovine herpesvirus 1 (BHV1) is the causative agent for infectious bovine rhinotracheitis and infectious pustular vulvovaginitis. After exposure to BHV1, antibodies against the virus can be detected using cattletype BHV1 gB Ab.

This product is a gB-blocking ELISA (enzyme-linked immunosorbent assay) with ready-to-use colored reagents. Samples are incubated with the test plate, which is coated with inactivated BHV1 antigen. If there are antibodies to glycoprotein B (gB) of the virus in the sample, they will bind to the antigen, thus blocking (inhibiting) the binding of an HRP-labeled, gB-specific monoclonal antibody. The blocking value (percentage of inhibition) can be calculated based on optical density measurements.

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.

There are two protocols for this product. They differ in the length of the incubation period. Serum and plasma samples are incubated for 2 hours with the test plate. For milk samples, an overnight incubation is needed. If antibodies to glycoprotein B of BHV1 are present, they will bind to the antigen on the plates. Unbound material is removed by rinsing.

The anti-gB-HRP conjugate is added. It contains an HRP-labeled, gB-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 gB Ab is for the detection of antibodies to glycoprotein B of bovine herpes virus 1 in serum, plasma, and milk samples from cattle.

Reference: Kramps, J.A. et al. (2004) Evaluation of tests for antibodies against bovine herpesvirus 1 performed in national reference laboratories in Europe. Vet. Microbiology 102 169

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