Cargando…
A reliable multiplex genotyping assay for HCV using a suspension bead array
The genotyping of the hepatitis C virus (HCV) plays an important role in the treatment of HCV because genotype determination has recently been incorporated into the treatment guidelines for HCV infections. Most current genotyping methods are unable to detect mixed genotypes from two or more HCV infe...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BlackWell Publishing Ltd
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321376/ https://www.ncbi.nlm.nih.gov/pubmed/25042084 http://dx.doi.org/10.1111/1751-7915.12140 |
_version_ | 1782356260412194816 |
---|---|
author | Yang, Yi-Chen Wang, Der-Yuan Cheng, Hwei-Fang Chuang, Eric Y Tsai, Mong-Hsun |
author_facet | Yang, Yi-Chen Wang, Der-Yuan Cheng, Hwei-Fang Chuang, Eric Y Tsai, Mong-Hsun |
author_sort | Yang, Yi-Chen |
collection | PubMed |
description | The genotyping of the hepatitis C virus (HCV) plays an important role in the treatment of HCV because genotype determination has recently been incorporated into the treatment guidelines for HCV infections. Most current genotyping methods are unable to detect mixed genotypes from two or more HCV infections. We therefore developed a multiplex genotyping assay to determine HCV genotypes using a bead array. Synthetic plasmids, genotype panels and standards were used to verify the target-specific primer (TSP) design in the assay, and the results indicated that discrimination efforts using 10 TSPs in a single reaction were extremely successful. Thirty-five specimens were then tested to evaluate the assay performance, and the results were highly consistent with those of direct sequencing, supporting the reliability of the assay. Moreover, the results from samples with mixed HCV genotypes revealed that the method is capable of detecting two different genotypes within a sample. Furthermore, the specificity evaluation results suggested that the assay could correctly identify HCV in HCV/human immunodeficiency virus (HIV) co-infected patients. This genotyping platform enables the simultaneous detection and identification of more than one genotype in a same sample and is able to test 96 samples simultaneously. It could therefore provide a rapid, efficient and reliable method of determining HCV genotypes in the future. |
format | Online Article Text |
id | pubmed-4321376 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BlackWell Publishing Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-43213762015-02-26 A reliable multiplex genotyping assay for HCV using a suspension bead array Yang, Yi-Chen Wang, Der-Yuan Cheng, Hwei-Fang Chuang, Eric Y Tsai, Mong-Hsun Microb Biotechnol Research Articles The genotyping of the hepatitis C virus (HCV) plays an important role in the treatment of HCV because genotype determination has recently been incorporated into the treatment guidelines for HCV infections. Most current genotyping methods are unable to detect mixed genotypes from two or more HCV infections. We therefore developed a multiplex genotyping assay to determine HCV genotypes using a bead array. Synthetic plasmids, genotype panels and standards were used to verify the target-specific primer (TSP) design in the assay, and the results indicated that discrimination efforts using 10 TSPs in a single reaction were extremely successful. Thirty-five specimens were then tested to evaluate the assay performance, and the results were highly consistent with those of direct sequencing, supporting the reliability of the assay. Moreover, the results from samples with mixed HCV genotypes revealed that the method is capable of detecting two different genotypes within a sample. Furthermore, the specificity evaluation results suggested that the assay could correctly identify HCV in HCV/human immunodeficiency virus (HIV) co-infected patients. This genotyping platform enables the simultaneous detection and identification of more than one genotype in a same sample and is able to test 96 samples simultaneously. It could therefore provide a rapid, efficient and reliable method of determining HCV genotypes in the future. BlackWell Publishing Ltd 2015-01 2014-07-10 /pmc/articles/PMC4321376/ /pubmed/25042084 http://dx.doi.org/10.1111/1751-7915.12140 Text en © 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology. http://creativecommons.org/licenses/by/3.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Yang, Yi-Chen Wang, Der-Yuan Cheng, Hwei-Fang Chuang, Eric Y Tsai, Mong-Hsun A reliable multiplex genotyping assay for HCV using a suspension bead array |
title | A reliable multiplex genotyping assay for HCV using a suspension bead array |
title_full | A reliable multiplex genotyping assay for HCV using a suspension bead array |
title_fullStr | A reliable multiplex genotyping assay for HCV using a suspension bead array |
title_full_unstemmed | A reliable multiplex genotyping assay for HCV using a suspension bead array |
title_short | A reliable multiplex genotyping assay for HCV using a suspension bead array |
title_sort | reliable multiplex genotyping assay for hcv using a suspension bead array |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321376/ https://www.ncbi.nlm.nih.gov/pubmed/25042084 http://dx.doi.org/10.1111/1751-7915.12140 |
work_keys_str_mv | AT yangyichen areliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT wangderyuan areliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT chenghweifang areliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT chuangericy areliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT tsaimonghsun areliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT yangyichen reliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT wangderyuan reliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT chenghweifang reliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT chuangericy reliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray AT tsaimonghsun reliablemultiplexgenotypingassayforhcvusingasuspensionbeadarray |