In situ hybridization and immunohistochemistry for the detection of porcine cytomegalovirus.
To establish in situ hybridization and immunohistochemistry based-assays for the detection of porcine cytomegalovirus, routinely processed renal tissue sections from 34 diseased piglets suspected of having the infection were obtained and examined. Using hematoxylin and eosin, porcine cytomegalovirus inclusion bodies were found in the nucleus of renal epithelial cells and capillary endothelial cells in the renal medulla in 30 cases. Inclusion bodies corresponding to porcine cytomegalovirus mRNA after in situ hybridization or porcine cytomegalovirus antigens after immunohistochemistry were easily determined. The cells were characterized by cytomegaly and basophilic intranuclear inclusion bodies. Using in situ hybridization, porcine cytomegalovirus mRNA were clearly detected in the nucleus and cytoplasm of the cells in 28 of the 30 (93.3%) cases. Using immunohistochemistry, porcine cytomegalovirus antigens were clearly detected in the cytoplasm of the cells in 21 of the 30 (70.0%) cases. Higher specificities and increased intensity of staining was observed with minimal background using in situ hybridization and immunohistochemistry compared with hematoxylin and eosin. Thus, the two established methods are useful and helpful tools for detecting the presence of a porcine cytomegalovirus infection.
Chuou Livestock Hygiene Service Center, 497 Iwatomi, Sakura, Chiba 285-0072, Japan.
Maki
M
Sekiguchi
Tomoyuki
T
Shibahara
Ayako
A
Miyazaki
Tomoko
T
Tajima
Shinya
S
Shimizu
Emmanuel
E
Kabali
Yoshiyuki
Y
Takano
Yosuke
Y
Sasaki
Masanori
M
Kubo
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D006651:Histocytochemistry / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D007668:Kidney / Q000821:virology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*
D000956:Antigens, Viral; D012333:RNA, Messenger; D012367:RNA, Viral