A fast and robust method for full genome sequencing of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Type 1 and Type 2
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A fast and robust method for full genome sequencing of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Type 1 and Type 2. / Kvisgaard, Lise K; Hjulsager, Charlotte K; Fahnøe, Ulrik; Breum, Solvej Ø; Ait-Ali, Tahar; Larsen, Lars E.
I: Journal of Virological Methods, Bind 193, Nr. 2, 11.2013, s. 697-705.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A fast and robust method for full genome sequencing of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Type 1 and Type 2
AU - Kvisgaard, Lise K
AU - Hjulsager, Charlotte K
AU - Fahnøe, Ulrik
AU - Breum, Solvej Ø
AU - Ait-Ali, Tahar
AU - Larsen, Lars E
N1 - Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.
PY - 2013/11
Y1 - 2013/11
N2 - PRRSV is a positive-sense RNA virus with a high degree of genetic variability among isolates. For diagnostic sensitivity and vaccine design it is essential to monitor PRRSV genetic diversity. However, to date only a few full genome sequences of PRRSV isolates have been made publicly available. In the present study, fast and robust methods for long range RT-PCR amplification and subsequent next generation sequencing (NGS) were developed and validated on nine Type 1 and nine Type 2 PRRSV viruses. The methods generated robust and reliable sequences both on primary material and cell culture adapted viruses and the protocols performed well on all three NGS platforms tested (Roche 454 FLX, Illumina HiSeq2000, and Ion Torrent PGM™ Sequencer). These methods will greatly facilitate the generation of more full genome PRRSV sequences globally.
AB - PRRSV is a positive-sense RNA virus with a high degree of genetic variability among isolates. For diagnostic sensitivity and vaccine design it is essential to monitor PRRSV genetic diversity. However, to date only a few full genome sequences of PRRSV isolates have been made publicly available. In the present study, fast and robust methods for long range RT-PCR amplification and subsequent next generation sequencing (NGS) were developed and validated on nine Type 1 and nine Type 2 PRRSV viruses. The methods generated robust and reliable sequences both on primary material and cell culture adapted viruses and the protocols performed well on all three NGS platforms tested (Roche 454 FLX, Illumina HiSeq2000, and Ion Torrent PGM™ Sequencer). These methods will greatly facilitate the generation of more full genome PRRSV sequences globally.
KW - Animals
KW - Genome, Viral
KW - High-Throughput Nucleotide Sequencing/methods
KW - Porcine Reproductive and Respiratory Syndrome/virology
KW - Porcine respiratory and reproductive syndrome virus/genetics
KW - RNA, Viral/genetics
KW - Reverse Transcriptase Polymerase Chain Reaction/methods
KW - Swine
KW - Virology/methods
U2 - 10.1016/j.jviromet.2013.07.019
DO - 10.1016/j.jviromet.2013.07.019
M3 - Journal article
C2 - 23891870
VL - 193
SP - 697
EP - 705
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 2
ER -
ID: 196714730