<abstract xmlns="http://www.w3.org/1999/xhtml"><bold>Introduction:</bold> The reverse transcription polymerase chain reaction (RT-PCR) is one of the most extensively used methods for identification of animals infected with bluetongue virus (BTV). There are several RT-PCR protocols published and several real-time RT-PCR (rtRT-PCR) commercial kits available on the market. Because Poland faced BTV-14 infection in 2012, different protocols were implemented in the country to confirm the RT-PCR results positive for this virus. The article presents a comparative study of several RT-PCR protocols and discusses their diagnostic reliability and applicability.<bold>Material and Methods:</bold> Six rtRT-PCR/RT-PCR protocols were compared for the laboratory diagnostic of fourteen BTV-14 isolates circulating in Poland in 2012–2014.<bold>Results:</bold> All 14 isolates were positive in the protocols of Shaw <italic>et al.</italic> (18), a commercial LSI NS3 kit, and Eschbaumer <italic>et al.</italic> (5). Four out of fourteen BTV-14 isolates gave positive results in Hoffmann’s 2 and 6 protocols and none of the 14 isolates yielded positive results in Maan <italic>et al.</italic> (8) method. Phylogenetic study of a short fragment of 450 nt of BTV segment 2 (258–696 positions) revealed 100% identity within Polish variants and with Russian and Spanish isolates.<bold>Conclusion:</bold> The paper points to the possible false negative results in the diagnosis of BTV infections depending on the protocol used.</abstract>

Introduction: The reverse transcription polymerase chain reaction (RT-PCR) is one of the most extensively used methods for identification of animals infected with bluetongue virus (BTV). There are several RT-PCR protocols published and several real-time RT-PCR (rtRT-PCR) commercial kits available on the market. Because Poland faced BTV-14 infection in 2012, different protocols were implemented in the country to confirm the RT-PCR results positive for this virus. The article presents a comparative study of several RT-PCR protocols and discusses their diagnostic reliability and applicability.Material and Methods: Six rtRT-PCR/RT-PCR protocols were compared for the laboratory diagnostic of fourteen BTV-14 isolates circulating in Poland in 2012–2014.Results: All 14 isolates were positive in the protocols of Shaw et al. (18), a commercial LSI NS3 kit, and Eschbaumer et al. (5). Four out of fourteen BTV-14 isolates gave positive results in Hoffmann’s 2 and 6 protocols and none of the 14 isolates yielded positive results in Maan et al. (8) method. Phylogenetic study of a short fragment of 450 nt of BTV segment 2 (258–696 positions) revealed 100% identity within Polish variants and with Russian and Spanish isolates.Conclusion: The paper points to the possible false negative results in the diagnosis of BTV infections depending on the protocol used.

Diagnostic reliability of different RT-PCR protocols for the detection of bluetongue virus serotype 14 (BTV-14)

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Introduction: The reverse transcription polymerase chain reaction (RT-PCR) is one of the most extensively used methods for identification of animals infected...

Diagnostic reliability of different RT-PCR protocols for the detection of bluetongue virus serotype 14 (BTV-14)

Published Online: Dec 27, 2017

Page range: 391 - 395

Received: Jul 11, 2017

Accepted: Dec 04, 2017

DOI: https://doi.org/10.1515/jvetres-2017-0065

Keywords
bluetongue, typing, genetic variability, Poland

© 2017 Anna Orłowska et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Diagnostic reliability of different RT-PCR protocols for the detection of bluetongue virus serotype 14 (BTV-14)

Published Online: Dec 27, 2017

Page range: 391 - 395

Received: Jul 11, 2017

Accepted: Dec 04, 2017

DOI: https://doi.org/10.1515/jvetres-2017-0065

Keywordsbluetongue, typing, genetic variability, Poland

© 2017 Anna Orłowska et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
bluetongue, typing, genetic variability, Poland