Detection of West Nile virus using formalin fixed paraffin embedded tissues in crows and horses: quantification of viral transcripts by real-time RT-PCR.
West Nile virus (WNV) RNA was quantified in WNV infected crows and horses with the help of a real-time reverse transcriptase-PCR assay. A 5' nuclease assay, based on NS5 gene detection with a fluorescent probe was used for quantifying WNV RNA using formalin fixed paraffin embedded tissue specimens. Quantitative detection of WNV RNA showed the presence of a higher amount of the viral RNA in crow tissues compared to equine tissues and these results correlated well with the detection of WNV antigen by immunostaining. In crows, the highest amount of virus was seen in the intestine and in horses in the brain.
Tewari
Deepanker
D
Pennsylvania Veterinary Laboratory, 2305 N Cameron St, Harrisburg, PA 17110, USA. dtewari@state.pa.us
Kim
Hyun
H
Feria
Willard
W
Russo
Brigite
B
Acland
Helen
H
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D001715:Bird Diseases / Q000821:virology; D005557:Formaldehyde; D006734:Horse Diseases / Q000821:virology; D006736:Horses / Q000821:virology*; D007150:Immunohistochemistry; D009928:Organ Specificity; D016612:Paraffin Embedding / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D020308:Songbirds / Q000821:virology*; D016707:Tissue Fixation; D014158:Transcription, Genetic; D014901:West Nile Fever / Q000662:veterinary / Q000821:virology; D014902:West Nile virus / Q000302:isolation & purification*
D000956:Antigens, Viral; D012367:RNA, Viral; D005557:Formaldehyde