Sheep poxvirus identification from clinical specimens by PCR, cell culture, immunofluorescence and agar gel immunoprecipitation assay

• Published in: Molecular and cellular probes Vol. 14; no. 5; pp. 305 - 310
• Main Authors: Mangana-Vougiouka, O, Markoulatos, P, Koptopoulos, G, Nomikou, K, Bakandritsos, N, Papadopoulos, P
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-10-2000
• Subjects: Animals; Biopsy; Cells, Cultured; Fluorescent Antibody Technique - methods; Polymerase Chain Reaction - methods; Poxviridae - genetics; Poxviridae - isolation & purification; Poxviridae Infections - virology; Precipitin Tests - methods; Sensitivity and Specificity; Sheep; Sheep Diseases - virology; sheep poxvirus, PCR, cell culture, DIFA, AGIPT; Skin - cytology; Skin - virology; sheep poxvirus, PCR, cell culture, DIFA, AGIPT
• ISSN: 0890-8508; 1096-1194
• DOI: 10.1006/mcpr.2000.0319

Some 40 clinical specimens of skin lesions from sheep pox suspected cases were investigated by four different diagnostic assays: PCR, virus isolation in lamb testis cell cultures, direct immunofluorescent assay (DIFA) and antigen detecting agar gel immune precipitation test (AGIPT). All the specimens were positive by PCR and virus isolation, 29 were positive by DIFA and 16 by AGIPT. Using virus isolation on cell cultures as the gold standard, the PCR sensitivity was 100%, while that of DIFA and AGIPT was 73% and 40%, respectively. Skin samples with orf lesions or normal skin biopsies were PCR-negative. Cross-reactions with orf virus were observed in three samples only in the AGIPT assay. The PCR described combines high specificity and sensitivity with speed. PCR was therefore shown to be the method of choice for sheep poxvirus diagnosis directly from clinical specimens.