Detection of bovine viral diarrhea virus in formalin fixed paraffin embedded tissue sections by real time RT-PCR (Taqman).
Real time quantitative RT-PCR (Taqman) identified specifically BVD virus in freshly processed formalin-fixed paraffin-embedded tissue sections and archival samples up to 7 years old. Samples included tissue from both acutely and persistently infected animals. To assess RNA degradation due to tissue handling and processing, freshly collected tissues from a calf persistently infected with BVD virus were stored at 4 degrees C or room temperature for up to 1 week prior to 24 h formalin fixation and routine histologic processing. Samples stored at 4 degrees C for up to a week prior to fixation were positive while samples stored at room temperature were positive at 74 h but became negative after 96 h. Fixation of fresh tissue in formalin for 1 week prior to processing resulted in a mild decrease in signal strength compared with tissue fixed for 24-48 h. Real time RT-PCR improves diagnosis of BVD infection by allowing prospective and retrospective identification of BVD virus in tissues processed routinely and stored for histologic evaluation.
Bhudevi
Bodreddigari
B
Animal Diagnostic Laboratory, Pennsylvania State University, University Park, PA 16802, USA. bbhudevi@criver.com
Weinstock
Daniel
D
D000818:Animals; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000821:virology; D002417:Cattle; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000302:isolation & purification*; D005404:Fixatives; D005557:Formaldehyde; D016612:Paraffin Embedding / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019914:Taq Polymerase / Q000378:metabolism; D016707:Tissue Fixation / Q000379:methods*; D014773:Virology / Q000379:methods
D005404:Fixatives; D012367:RNA, Viral; D005557:Formaldehyde; D019914:Taq Polymerase