scholarly journals Optimization of In situ Hybridization Protocols for Detection of Feline Herpesvirus 1

2003 ◽  
Vol 65 (9) ◽  
pp. 1031-1032 ◽  
Author(s):  
Okjin KIM ◽  
Seong Joon YI
2005 ◽  
Vol 69 (3) ◽  
pp. 345-349 ◽  
Author(s):  
José Vassallo ◽  
Leandro Azevedo de Camargo ◽  
Cristiano Aparecido Chagas ◽  
Glauce Aparecida Pinto ◽  
Luiza Hayashi Endo

1989 ◽  
Vol 1 (1) ◽  
pp. 34-38 ◽  
Author(s):  
Kenny V. Brock ◽  
Leon N. D Potgieter

The use of biotinylated DNA hybridization probes for clinical detection of bovine herpesvirus-1 was investigated. Biotinylated DNA hybridization probes were prepared from bovine herpesvirus-1 DNA purified from infected cell cultures. The viral DNA was nick translated in the presence of biotin-dUTP with DNA polymerase to incorporate biotin into the newly synthesized strand. The probe was tested for specificity in in situ hybridization assays with bovine herpesvirus-1 DNA. Hybridization was detected using avidin-fluorescein single sandwich systems and an avidin-globulin with anti-globulin-fluorescein double sandwich system. Hybridization was detected by specific fluorescence of infected cells. Fluorescence was present only in bovine herpesvirus-1-infected cell culture and not in noninfected cell culture or cell cultures infected with several other viruses. The assay was performed in 6 hr.


2000 ◽  
Vol 12 (2) ◽  
pp. 186-191 ◽  
Author(s):  
A. Suchy ◽  
B. Bauder ◽  
W. Gelbmann ◽  
C. V. Löhr ◽  
J. P. Teifke ◽  
...  

An adult domestic shorthair cat had severe chemosis due to purulent and necrotizing blepharitis and conjunctivitis. Purulent rhinitis, necrotizing glossitis, and dermatitis were also diagnosed. The cat was positive for feline immundeficiency virus and feline leukemia virus. Histologically, intranuclear Cowdry type A inclusions were found within numerous epithelial cells adjacent to the lesions in skin, conjunctiva, and tongue. Electron microscopic examination revealed herpesviral particles within the lesions. Paraffin-embedded skin and tongue tissues were processed in a polymerase chain reaction, using primers to amplify a 306-bp region of the thymidine kinase gene of feline herpesvirus type 1, resulting in a distinct amplification product of the predicted size. The distribution of feline herpesvirus was demonstrated by immunohistochemistry and nonradioactive in situ hybridization. Positive immunostaining was found in nuclei and cytoplasm of numerous epithelial cells within and next to the lesions, whereas in situ hybridization, performed with a digoxigenin-labeled double-stranded DNA probe, revealed hybridization signal only in nuclei of intact epithelial cells. Neither immunohistochemistry nor in situ hybridization showed feline herpesvirus type 1 in tissues of lungs, liver, spleen, intestine, or brain.


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