Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus

The emergence in Japan of field isolates of type 1 porcine reproductive and respiratory syndrome virus (PRRSV) suggests problems with control. We therefore developed a one-step real-time reverse transcription polymerase chain reaction (qRT-PCR) with improved sensitivity that detects as little as 1 ×...

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Autores principales: ISEKI, Hiroshi, TAKAGI, Michihiro, KURODA, Yoshiko, KATSUDA, Ken, MIKAMI, Osamu, TSUNEMITSU, Hiroshi, YAMAKAWA, Makoto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2014
Materias:
Virology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4221178/
https://www.ncbi.nlm.nih.gov/pubmed/25047905
http://dx.doi.org/10.1292/jvms.14-0010
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author ISEKI, Hiroshi
TAKAGI, Michihiro
KURODA, Yoshiko
KATSUDA, Ken
MIKAMI, Osamu
TSUNEMITSU, Hiroshi
YAMAKAWA, Makoto
author_facet ISEKI, Hiroshi
TAKAGI, Michihiro
KURODA, Yoshiko
KATSUDA, Ken
MIKAMI, Osamu
TSUNEMITSU, Hiroshi
YAMAKAWA, Makoto
author_sort ISEKI, Hiroshi
collection PubMed
description The emergence in Japan of field isolates of type 1 porcine reproductive and respiratory syndrome virus (PRRSV) suggests problems with control. We therefore developed a one-step real-time reverse transcription polymerase chain reaction (qRT-PCR) with improved sensitivity that detects as little as 1 × 10(−2) TCID(50)/ml of viral RNA. We tested serum samples collected in January and September 2008, October 2009 and January 2011 from a farm with an outbreak and found infected pigs between January and September 2008, but not in January 2011. Further, between 2008 and 2011, we did not detect infection in pigs at 8 nearby farms or in 2,052 serum samples collected from pigs from 74 farms in 12 prefectures. This assay should help prevent future outbreaks.
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spelling pubmed-42211782014-11-06 Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus ISEKI, Hiroshi TAKAGI, Michihiro KURODA, Yoshiko KATSUDA, Ken MIKAMI, Osamu TSUNEMITSU, Hiroshi YAMAKAWA, Makoto J Vet Med Sci Virology The emergence in Japan of field isolates of type 1 porcine reproductive and respiratory syndrome virus (PRRSV) suggests problems with control. We therefore developed a one-step real-time reverse transcription polymerase chain reaction (qRT-PCR) with improved sensitivity that detects as little as 1 × 10(−2) TCID(50)/ml of viral RNA. We tested serum samples collected in January and September 2008, October 2009 and January 2011 from a farm with an outbreak and found infected pigs between January and September 2008, but not in January 2011. Further, between 2008 and 2011, we did not detect infection in pigs at 8 nearby farms or in 2,052 serum samples collected from pigs from 74 farms in 12 prefectures. This assay should help prevent future outbreaks. The Japanese Society of Veterinary Science 2014-07-18 2014-10 /pmc/articles/PMC4221178/ /pubmed/25047905 http://dx.doi.org/10.1292/jvms.14-0010 Text en ©2014 The Japanese Society of Veterinary Science http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Virology
ISEKI, Hiroshi
TAKAGI, Michihiro
KURODA, Yoshiko
KATSUDA, Ken
MIKAMI, Osamu
TSUNEMITSU, Hiroshi
YAMAKAWA, Makoto
Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title_full Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title_fullStr Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title_full_unstemmed Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title_short Application of a SYBR® Green One Step Real-time RT-PCR Assay to Detect Type 1 Porcine Reproductive and Respiratory Syndrome Virus
title_sort application of a sybr® green one step real-time rt-pcr assay to detect type 1 porcine reproductive and respiratory syndrome virus
topic Virology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4221178/
https://www.ncbi.nlm.nih.gov/pubmed/25047905
http://dx.doi.org/10.1292/jvms.14-0010
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