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Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids
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Molecular and immunological detection of bovine herpesvirus-1 in clinical specimens

Alaa A El-Kholy1

  • 1Veterinary Serum & Vaccine Research Institute, Abbasia, Cairo, Egypt.

The Egyptian Journal of Immunology
|November 6, 2007
PubMed

Insights

A new nested PCR method effectively detects bovine herpesvirus-1 (BHV-1) in cattle and buffalo, outperforming traditional methods. This sensitive assay is crucial for controlling BHV-1 infections in animals.

Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • Animal Health

Background:

  • Bovine herpesvirus-1 (BHV-1) causes significant economic losses in cattle, buffalo, and sheep.
  • Accurate and sensitive diagnostic tools are essential for effective BHV-1 control programs.

Purpose of the Study:

  • To evaluate the efficacy of a nested polymerase chain reaction (nested-PCR) targeting the glycoprotein B (gB) gene for BHV-1 detection.
  • To compare the performance of nested-PCR with traditional methods like virus isolation (VI), indirect immunofluorescence (IFA), and In situ immuno-peroxidase (IPr) staining.

Main Methods:

  • Nested-PCR was developed using the BHV-1 gB gene as a target.
  • Clinical specimens (nasal swabs, buffy coats, sera) from cattle, buffalo, and sheep in Egypt were tested.
  • Results were compared with VI, IFA, and IPr staining.

Main Results:

  • Nested-PCR detected BHV-1 in 16.1% of cattle and 7.3% of buffalo specimens.
  • Nested-PCR demonstrated higher sensitivity and discriminative power than immunodetection methods.
  • BHV-1 viral DNA was detected directly from cattle and buffalo specimens with 100% accuracy using nested-PCR.
  • Sheep isolates showed discrepancies, with some positive by IFA/IPr/first-round PCR but negative by nested-PCR, suggesting other related herpesviruses.

Conclusions:

  • The BHV-1 gB-based nested-PCR is a highly sensitive, discriminative, and rapid tool for BHV-1 diagnosis.
  • This method is recommended for epidemiological studies and control programs, including routine screening of animals and biologics.
  • The nested-PCR assay can differentiate BHV-1 from other ruminant herpesviruses, preventing diagnostic confusion.

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