Development of a semi-nested reverse transcription polymerase chain reaction assay for the retrospective diagnosis of canine distemper virus infection.
Formalin-fixed, paraffin-embedded tissue blocks from 14 dogs were used to test the utility of a newly developed semi-nested reverse transcription polymerase chain reaction (RT-PCR) assay for canine distemper virus (CDV). The results from this new test were compared with those of histopathologic examination, fluorescent antibody detection (FA), and immunohistochemistry (IHC). The semi-nested RT-PCR protocol was used to detect CDV RNA in 9 of the 10 cases that were positive by at least 1 of the immunologic (FA and IHC) methods. Sequence data indicated that the amplified strains of CDV are more closely related to a naturally occurring strain than to a vaccine strain. Thus, the semi-nested RT-PCR assay is a useful diagnostic method applicable to the retrospective diagnosis of CDV infection. Sequence determination may yield molecular epidemiologic information regarding vaccine efficacy.
Stanton
James B
JB
Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
Poet
Steven
S
Frasca
Salvatore
S
Bienzle
Dorothee
D
Brown
Corrie C
CC
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000473:pathology / Q000821:virology; D004216:Distemper / Q000175:diagnosis* / Q000473:pathology / Q000821:virology; D004217:Distemper Virus, Canine / Q000235:genetics* / Q000302:isolation & purification*; D004285:Dogs; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D012367:RNA, Viral