11-Title: Molecular detection of ovine herpes virus-2 in a buffaloe by targeting ORF63 gene using sybr green real time PCR

Authors: T Nagendra Reddy, M Nagaraja  and A Phanindra Reddy

Source: Ruminant Science (2022)-11(2):309-312.

How to cite this manuscript: Reddy T Nagendra, Nagaraja M and Reddy A Phanindra (2022). Molecular detection of ovine herpes virus-2 in a buffaloe by targeting ORF63 gene using sybr green real time PCR. Ruminant Science 11(2):309-312.

Abstract

A buffalo was presented with a history of corneal opacity, fever and anorexia. The clinical examination revealed high rectal temperature (106ÚF), bilateral corneal opacity, lacrimal discharges, mucopurulent nasal discharge and crust formation in the nostrils. Based on clinical signs the clinical case was suspected for Malignant catarrhal fever (MCF). Hence this work was aimed at detection of ovine herpes virus-2 from samples using SYBR Green Real time PCR assay. The nasal swabs, ocular swabs and blood in EDTA samples were collected from ailing buffalo. The DNA was isolated from the samples by using Trizol method. In the present study, molecular detection was done by targeting the ORF63 gene using SYBR Green Real time PCR. The clear-cut ORF63 gene amplification was observed and recorded the amplification plot and melt curve. The sample was found to be positive at ct value 25 and the product Tm was 75.5. All the samples that were collected from ailing buffalo, including the nasal swabs, ocular swabs and blood in EDTA were screened for MCF and found positive reactions by Optimised SYBR Green Real Time PCR.

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