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Related Experiment Videos

Detection of bovine herpesvirus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk.

G J Wellenberg1, E R Verstraten, S Belák

  • 1Division Infectious Diseases and Food Chain Quality, Institute for Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB, Lelystad, The Netherlands.

Journal of Virological Methods
|August 3, 2001
PubMed
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Developed polymerase chain reaction (PCR) assays for bovine herpesvirus 4 (BHV4) glycoprotein B (gB) and thymidine kinase (TK) DNA in milk. These PCR methods offer superior sensitivity and reliability for detecting BHV4 infections in cattle.

Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • Bovine Infectious Diseases

Background:

  • Bovine herpesvirus 4 (BHV4) is an important pathogen affecting cattle health.
  • Accurate and sensitive detection methods for BHV4 in milk are crucial for disease control.
  • Existing diagnostic methods like virus isolation have limitations in sensitivity and speed.

Purpose of the Study:

  • To develop and validate novel polymerase chain reaction (PCR) assays for the detection of BHV4 DNA in bovine milk.
  • To compare the sensitivity and specificity of the developed PCR assays against traditional methods like virus isolation.
  • To assess the utility of these assays for routine screening of BHV4 in dairy herds.

Main Methods:

  • Development of a PCR assay for BHV4 glycoprotein B (gB) DNA detection.

Related Experiment Videos

  • Modification of a nested-PCR assay for BHV4 thymidine kinase (TK) DNA detection.
  • Inclusion of internal control templates to identify false-negative PCR results.
  • Validation of specificity, sensitivity, and reproducibility using BHV4 strains and unrelated viruses.
  • Comparison of PCR assay performance with virus isolation in naturally infected milk samples.

    • Main Results:

    • Both gB-PCR and TK-PCR assays demonstrated high specificity, correctly identifying all 31 BHV4 strains and none of the 14 unrelated viruses.
    • The TK-PCR assay detected as few as 1-3 copies of BHV4 DNA, while the gB-PCR detected 2-10 copies.
    • BHV4 DNA was detected in 93% (gB-PCR) and 95% (TK-PCR) of infected milk samples, significantly higher than virus isolation (61%).
    • The gB-PCR and TK-PCR assays were 10-fold and 55-fold more sensitive, respectively, than virus isolation.
    • BHV4 DNA was detected in milk from cows with clinical mastitis, indicating the assays' relevance in disease outbreaks.

    Conclusions:

    • The developed PCR assays are rapid, reliable, and highly sensitive for detecting BHV4 DNA in bovine milk.
    • The gB-PCR assay is recommended for screening large volumes of milk samples due to its lower labor requirements.
    • These molecular diagnostic tools can significantly improve the surveillance and control of BHV4 infections in cattle herds.