Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction.

Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Research Abstract Details 

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Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Abstract Text:

A D Gruber,V Moennig,M Hewicker-Trautwein,G Trautwein,

Detection of DNA or RNA in formalin-fixed, paraffin-embedded tissues using polymerase chain reaction (PCR) may be hindered by degradation of nucleic acids during tissue collection, preparation and archivation. This study describes investigations on the effect of formalin fixation and prolonged storage of paraffin-embedded tissues on bovine viral-diarrhoea (BVD)-virus RNA as a model system. Brain tissues from eight persistently BVDV-infected calves containing high amounts of the virus were fixed in 5% neutral-buffered formalin or 10% non-buffered formalin for different fixation times, respectively, and paraffin embedded. Subsequent detection of an 803 bp fragment from single tissue sections using nested PCR after reverse transcription (nested RT-PCR) demonstrated a loss of detectability of viral RNA after more than 10 days (10% non-buffered formalin) and 3 months (5% neutral-buffered formalin) of fixation. Additional studies with 280 initially BVDV-positive brain tissues from 25 persistently BVDV-infected calves after storage of up to 10 years revealed a loss of detectable RNA after more than 1 year of storage. For estimation of the higher sensitivity of nested RT-PCR compared to single step RT-PCR, serially diluted BVD virus suspensions were examined using both methods. Nested RT-PCR was found to be about 100-fold more sensitive than single-step RT-PCR, and is therefore recommended as the appropriate technique for archival studies.

Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Publishing Authors By Initials

AD Gruber,V Moennig,M Hewicker-Trautwein,G Trautwein,

For similar diagnosis: laboratory techniques and procedures: specimen handling: preservation, biological: tissue preservation research abstracts see: diagnosis: laboratory techniques and procedures: specimen handling: preservation, biological: tissue preservation research

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Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Zentralblatt für Veterinärmedizin. Reihe B. Journa

VOLUME: 41

Page Numbers: 654-61

Journal Abbreviation: Zentralblatt Veterinarmedizin

ISSN: 0514-7166

DAY: 13

MONTH: Dec

YEAR: 1994

Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 331325

Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Keywords Mesh Terms:

KEYWORDS: Tissue Preservation

MESH TERMS: veterinary

Chemical & Substance for Abstract: Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction. Information

Substance Name: Formaldehyde

Registry Number: 50-00-0

Grant and Affiliation Information for Effect of formalin fixation and long-term storage on the detectability of bovine viral-diarrhoea-virus (BVDV) RNA in archival brain tissue using polymerase chain reaction.

AFFILIATION: Institute of Pathology, School of Veterinary Medicine, Hannover, Germany.

Country: GERMANY

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MEDLINETA: Zentralbl Veterinarmed B

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