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Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host

To date, recombination between different strains of the avian alphaherpesvirus infectious laryngotracheitis virus (ILTV) has only been detected in field samples using full genome sequencing and sequence analysis. These previous studies have revealed that natural recombination is widespread in ILTV a...

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Autores principales: Loncoman, Carlos A., Hartley, Carol A., Coppo, Mauricio J. C., Vaz, Paola K., Diaz-Méndez, Andrés, Browning, Glenn F., Lee, Sang-won, Devlin, Joanne M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5370143/
https://www.ncbi.nlm.nih.gov/pubmed/28350819
http://dx.doi.org/10.1371/journal.pone.0174590
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author Loncoman, Carlos A.
Hartley, Carol A.
Coppo, Mauricio J. C.
Vaz, Paola K.
Diaz-Méndez, Andrés
Browning, Glenn F.
Lee, Sang-won
Devlin, Joanne M.
author_facet Loncoman, Carlos A.
Hartley, Carol A.
Coppo, Mauricio J. C.
Vaz, Paola K.
Diaz-Méndez, Andrés
Browning, Glenn F.
Lee, Sang-won
Devlin, Joanne M.
author_sort Loncoman, Carlos A.
collection PubMed
description To date, recombination between different strains of the avian alphaherpesvirus infectious laryngotracheitis virus (ILTV) has only been detected in field samples using full genome sequencing and sequence analysis. These previous studies have revealed that natural recombination is widespread in ILTV and have demonstrated that recombination between two attenuated ILTV vaccine strains generated highly virulent viruses that produced widespread disease within poultry flocks in Australia. In order to better understand ILTV recombination, this study developed a TaqMan single nucleotide polymorphism (SNP) genotyping assay to detect recombination between two field strains of ILTV (CSW-1 and V1-99 ILTV) under experimental conditions. Following in vivo co-inoculation of these two ILTV strains in specific pathogen free (SPF) chickens, recovered viruses were plaque purified and subjected to the SNP genotyping assay. This assay revealed ILTV recombinants in all co-inoculated chickens. In total 64/87 (74%) of the recovered viruses were recombinants and 23 different recombination patterns were detected, with some of them occurring more frequently than others. The results from this study demonstrate that the TaqMan SNP genotyping assay is a useful tool to study recombination in ILTV and also show that recombination occurs frequently during experimental co-infection with ILTV in SPF chickens. This tool, when used to assess ILTV recombination in the natural host, has the potential to greatly contribute to our understanding of alphaherpesvirus recombination.
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spelling pubmed-53701432017-04-06 Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host Loncoman, Carlos A. Hartley, Carol A. Coppo, Mauricio J. C. Vaz, Paola K. Diaz-Méndez, Andrés Browning, Glenn F. Lee, Sang-won Devlin, Joanne M. PLoS One Research Article To date, recombination between different strains of the avian alphaherpesvirus infectious laryngotracheitis virus (ILTV) has only been detected in field samples using full genome sequencing and sequence analysis. These previous studies have revealed that natural recombination is widespread in ILTV and have demonstrated that recombination between two attenuated ILTV vaccine strains generated highly virulent viruses that produced widespread disease within poultry flocks in Australia. In order to better understand ILTV recombination, this study developed a TaqMan single nucleotide polymorphism (SNP) genotyping assay to detect recombination between two field strains of ILTV (CSW-1 and V1-99 ILTV) under experimental conditions. Following in vivo co-inoculation of these two ILTV strains in specific pathogen free (SPF) chickens, recovered viruses were plaque purified and subjected to the SNP genotyping assay. This assay revealed ILTV recombinants in all co-inoculated chickens. In total 64/87 (74%) of the recovered viruses were recombinants and 23 different recombination patterns were detected, with some of them occurring more frequently than others. The results from this study demonstrate that the TaqMan SNP genotyping assay is a useful tool to study recombination in ILTV and also show that recombination occurs frequently during experimental co-infection with ILTV in SPF chickens. This tool, when used to assess ILTV recombination in the natural host, has the potential to greatly contribute to our understanding of alphaherpesvirus recombination. Public Library of Science 2017-03-28 /pmc/articles/PMC5370143/ /pubmed/28350819 http://dx.doi.org/10.1371/journal.pone.0174590 Text en © 2017 Loncoman et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Loncoman, Carlos A.
Hartley, Carol A.
Coppo, Mauricio J. C.
Vaz, Paola K.
Diaz-Méndez, Andrés
Browning, Glenn F.
Lee, Sang-won
Devlin, Joanne M.
Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title_full Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title_fullStr Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title_full_unstemmed Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title_short Development and application of a TaqMan single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
title_sort development and application of a taqman single nucleotide polymorphism genotyping assay to study infectious laryngotracheitis virus recombination in the natural host
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5370143/
https://www.ncbi.nlm.nih.gov/pubmed/28350819
http://dx.doi.org/10.1371/journal.pone.0174590
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