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

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.


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.


Fernandez-Aguilar X, Esperon F, Cabezon O, Velarde R, Mentaberre G, Delicado V, Lopez-Olvera JR (2016). Identification of a herpesvirus belonging to the malignant catarrhal fever group of viruses in Pyrenean chamois (Rupicapra p pyrenaica). Archives of Virology 161(11):3249-3253.

Galbraith J, Rodas-Gonzalez A, Lopez-Campos O, Juarez M and Aalhus J (2014). Bison meat: Characteristics, challenges and opportunities. Animal Frontiers 4(4):68-73.

Headley SA, Pimentel LA, Oliveira VHS, Toma HS, Alfieri AF, Carvalho AM and Alfieri AA (2015). Transplacental transmission of ovine herpesvirus 2 in cattle with sheep-associated malignant catarrhal fever. Journal of Comparative Pathology 153(4):206-211.

Headley SA, de Oliveira TES and Cunha CW (2020). A review of the epidemiological, clinical, and pathological aspects of malignant catarrhal fever in Brazil. Brazilian Journal of Microbiology 51:1405-1432.

Jarquin R, Schultz J, Hanning I and Ricke SC (2009). Development of a real-time polymerase chain reaction assay for the simultaneous detection of Mycoplasma gallisepticum and Mycoplasma synoviae under industry conditions. Avian Diseases 53(1):73-77

Kiss I, Matiz K, Kaszanyitzky É, Chávez Y and Johansson KE (1997). Detection and identification of avian mycoplasmas by polymerase chain reaction and restriction fragment length polymorphism assay. Veterinary Microbiology 58(1):23-30.

Li X, Xie W, Xie C, Huang C, Zhu J, Liang Z and Zhong C (2014). Curcumin modulates miR 19/PTEN/AKT/p53 axis to suppress bisphenol A induced MCF 7 breast cancer cell proliferation. Phytotherapy Research 28(10):1553-1560

Liggitt HD and DeMartini JC (1980). The pathomorphology of malignant catarrhal fever: II multisystemic epithelial lesions. Veterinary Pathology 17(1):73-83.

Moore DA, Kohrs P, Baszler T, Faux C, Sathre P, Wenz JR and Li H (2010). Outbreak of malignant catarrhal fever among cattle associated with a state livestock exhibition. Journal of the American Veterinary Medical Association 237(1):87-92.

Phillips G, Lopman B, Tam CC, Iturriza-Gomara M, Brown D and Gray J (2009). Diagnosing norovirus-associated infectious intestinal disease using viral load. BMC Infectious Diseases 9(1):1-9.

Sailo L and Das R (2016). Heat stress in livestock: Impacts and ameliorative strategies-A Review. International Journal of Bio-Resource and Stress Management 7:1.

Shandiz SAS, Khosravani M, Mohammadi S, Noorbazargan H, Mirzaie A, Inanlou DN and Keshavarz-Pakseresht B (2016). Evaluation of imatinib mesylate (Gleevec) on KAI1/CD82 gene expression in breast cancer MCF-7 cells using quantitative real-time PCR. Asian Pacific Journal of Tropical Biomedicine 6(2):159-163.

Simon S, Li H, O’Toole D, Crawford TB and Oaks JL (2003). The vascular lesions of a cow and bison with sheep-associated malignant catarrhal fever contain ovine herpesvirus 2-infected CD8+ T lymphocytes. Journal of General Virology 84(8):2009-2013.

Sood R, Khandia R, Bhatia S, Hemadri D, Kumar M, Patil SS and Kulkarni DD (2014). Detection and molecular characterization of naturally transmitted sheep associated malignant catarrhal fever in cattle in India. Tropical Animal Health and Production 46(6):1037-1043.

Sprygin AV, Andreychuk DB, Kolotilov AN, Volkov MS, Runina IA, Mudrak NS and Perevozchikova NA (2010). Development of a duplex real-time TaqMan PCR assay with an internal control for the detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from commercial and backyard poultry. Avian Pathology 39(2):99-109.

Stahel AB, Baggenstos R, Engels M, Friess M and Ackermann M (2013). Two different macaviruses, ovine herpesvirus-2 and caprine herpesvirus-2, behave differently in water buffaloes than in cattle or in their respective reservoir species. PloS one 8(12):e83695

Wambua L, Wambua PN, Ramogo AM, Mijele D and Otiende MY (2016). Wildebeest-associated malignant catarrhal fever: Perspectives for integrated control of a lymphoproliferative disease of cattle in sub-Saharan Africa. Archives of Virology 161(1):1-10.