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Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing
Background and objectives: The human respiratory syncytial virus (hRSV) is among the important respiratory pathogens affecting children. Genotype-specific attachment (G) gene sequencing is usually used to determine the virus genotype. The reliability of the fusion (F) gene vs. G gene genotype-specif...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6571984/ https://www.ncbi.nlm.nih.gov/pubmed/31137571 http://dx.doi.org/10.3390/medicina55050169 |
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author | Al Aboud, Daifullah Al Aboud, Nora M. Al-Malky, Mater I. R. Abdel-Moneim, Ahmed S. |
author_facet | Al Aboud, Daifullah Al Aboud, Nora M. Al-Malky, Mater I. R. Abdel-Moneim, Ahmed S. |
author_sort | Al Aboud, Daifullah |
collection | PubMed |
description | Background and objectives: The human respiratory syncytial virus (hRSV) is among the important respiratory pathogens affecting children. Genotype-specific attachment (G) gene sequencing is usually used to determine the virus genotype. The reliability of the fusion (F) gene vs. G gene genotype-specific sequencing was screened. Materials and Methods: Archival RNA from Saudi children who tested positive for hRSV-A were used. Samples were subjected to a conventional one-step RT-PCR for both F and G genes and direct gene sequencing of the amplicons using the same primer sets. Phylogeny and mutational analysis of the obtained sequences were conducted. Results: The generic primer set succeeded to amplify target gene sequences. The phylogenetic tree based on partial F gene sequencing resulted in an efficient genotyping of hRSV-A strains equivalent to the partial G gene genotyping method. NA1, ON1, and GA5 genotypes were detected in the clinical samples. The latter was detected for the first time in Saudi Arabia. Different mutations in both conserved and escape-mutant domains were detected in both F and G. Conclusion: It was concluded that a partial F gene sequence can be used efficiently for hRSV-A genotyping. |
format | Online Article Text |
id | pubmed-6571984 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-65719842019-06-18 Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing Al Aboud, Daifullah Al Aboud, Nora M. Al-Malky, Mater I. R. Abdel-Moneim, Ahmed S. Medicina (Kaunas) Article Background and objectives: The human respiratory syncytial virus (hRSV) is among the important respiratory pathogens affecting children. Genotype-specific attachment (G) gene sequencing is usually used to determine the virus genotype. The reliability of the fusion (F) gene vs. G gene genotype-specific sequencing was screened. Materials and Methods: Archival RNA from Saudi children who tested positive for hRSV-A were used. Samples were subjected to a conventional one-step RT-PCR for both F and G genes and direct gene sequencing of the amplicons using the same primer sets. Phylogeny and mutational analysis of the obtained sequences were conducted. Results: The generic primer set succeeded to amplify target gene sequences. The phylogenetic tree based on partial F gene sequencing resulted in an efficient genotyping of hRSV-A strains equivalent to the partial G gene genotyping method. NA1, ON1, and GA5 genotypes were detected in the clinical samples. The latter was detected for the first time in Saudi Arabia. Different mutations in both conserved and escape-mutant domains were detected in both F and G. Conclusion: It was concluded that a partial F gene sequence can be used efficiently for hRSV-A genotyping. MDPI 2019-05-20 /pmc/articles/PMC6571984/ /pubmed/31137571 http://dx.doi.org/10.3390/medicina55050169 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Al Aboud, Daifullah Al Aboud, Nora M. Al-Malky, Mater I. R. Abdel-Moneim, Ahmed S. Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title | Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title_full | Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title_fullStr | Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title_full_unstemmed | Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title_short | Genotyping of Type A Human Respiratory Syncytial Virus Based on Direct F Gene Sequencing |
title_sort | genotyping of type a human respiratory syncytial virus based on direct f gene sequencing |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6571984/ https://www.ncbi.nlm.nih.gov/pubmed/31137571 http://dx.doi.org/10.3390/medicina55050169 |
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