Procedures for reproducible detection of rabies virus antigen mRNA and genome in situ in formalin-fixed tissues

Procedures allowing the reproducible in situ detection of rabies virus antigen and RNAs (both genome and message) in formalin-fixed tissue are described. These procedures can be used on sequential tissue sections and thereby permit comparison of results from tests detecting both antigen and RNA in t...

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Bibliographic Details
Published in:Journal of virological methods 1997-08, Vol.67 (1), p.5-12
Main Authors: Warner, Cynthia K, Whitfield, Sylvia G, Fekadu, Makonnen, Ho, Hue
Format: Article
Language:English
Subjects:
Animals
Antigen
Antigens, Viral - analysis
Antigens, Viral - genetics
Biological and medical sciences
Brain - virology
Chiroptera - virology
Endopeptidase K
Fixatives
Fluorescent Antibody Technique
Formaldehyde
Formalin-fixed detection
Fundamental and applied biological sciences. Psychology
Genome
In Situ Hybridization - methods
Mice
Microbiology
Molecular Sequence Data
mRNA
Rabies
Rabies - diagnosis
rabies virus
Rabies virus - genetics
Rabies virus - immunology
Rabies virus - isolation & purification
Rabies-related
RNA, Messenger - analysis
RNA, Viral - analysis
Techniques used in virology
Tissue Preservation
Virology
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Summary:Procedures allowing the reproducible in situ detection of rabies virus antigen and RNAs (both genome and message) in formalin-fixed tissue are described. These procedures can be used on sequential tissue sections and thereby permit comparison of results from tests detecting both antigen and RNA in the same tissue. This antigen-detecting procedure has also been used to identify both the phylogenetically distant rabies viruses from silver-haired bat and vampire bat and the rabies-related viruses Mokola, Duvenhage, and Lagos bat. One of the critical steps in these procedures is the digestion (and the resulting exposure of the target molecules) with proteinase K. These methods may be useful for the identification of other viruses of public health importance. Because in many situations only formalin-fixed tissue is available for postmortem diagnosis, the technical ability to identify a virus antigen and nucleic acid in such tissues greatly extends potential diagnostic capabilities.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(97)00068-2