Double in situ hybridization for simultaneous detection and differentiation of porcine circovirus 1 and 2 in pigs with postweaning multisystemic wasting syndrome.
Double in situ hybridization using a digoxigenin-labelled porcine circovirus 1 (PCV1) and biotinylated PCV2 probe, was developed for the simultaneous detection and differentiation of PCV1 and PCV2 in formalin-fixed, paraffin-embedded tissues from pigs with postweaning multisystemic wasting syndrome. The combination of an alkaline phosphatase conjugated antidigoxigenin system with alkaline phosphatase conjugated streptavidin-biotin system allowed identification of PCV1 and/or PCV2. No evidence of cross-reaction was observed. Positive cells exhibited a red or dark brown reaction product for PCV1 and PCV2, respectively. Both PCV DNAs were observed mainly in the cytoplasm but occasionally in the nucleus. Co-localization of hybridization signal for both PCV1 and PCV2 was present in macrophages and multinucleated giant cells of the lymph node and spleen. This double-labelling technique for the differentiation between PCV1 and PCV2 is suitable for pathogenesis studies and diagnostic applications.
Kim
J
J
Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Kyounggi-Do, Republic of Korea.
Chae
C
C
D000818:Animals; D018173:Circoviridae Infections / Q000150:complications* / Q000662:veterinary*; D017925:Circovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization; D008198:Lymph Nodes / Q000821:virology; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D013154:Spleen / Q000821:virology; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000821:virology*; D019282:Wasting Syndrome / Q000150:complications* / Q000662:veterinary*; D014886:Weaning
D012367:RNA, Viral