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A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses()
A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of California (CAL) serogroup viruses and Cache Valley virus (CVV), for use in human surveillance. The targets selected for the assay were the sequences encoding the nucleocapsid pro...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Elsevier Inc.
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774246/ https://www.ncbi.nlm.nih.gov/pubmed/19748425 http://dx.doi.org/10.1016/j.diagmicrobio.2009.07.001 |
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author | Wang, Heng Nattanmai, Seela Kramer, Laura D. Bernard, Kristen A. Tavakoli, Norma P. |
author_facet | Wang, Heng Nattanmai, Seela Kramer, Laura D. Bernard, Kristen A. Tavakoli, Norma P. |
author_sort | Wang, Heng |
collection | PubMed |
description | A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of California (CAL) serogroup viruses and Cache Valley virus (CVV), for use in human surveillance. The targets selected for the assay were the sequences encoding the nucleocapsid protein of CAL and the G1 glycoprotein of CVV. Conserved regions were selected by aligning genetic sequences from various strains available in the GenBank database. Primers and probes were selected in conserved regions. The assay sensitivity was 75 gene copies (gc)/reaction for CAL serogroup viruses and 30 gc/reaction for CVV. The performance of the assay was linear over at least 6 log(10) gc. The assay was specific, given that it did not cross-react with a variety of pathogens. It did, however, detect 11 viruses within the CAL serogroup and 12 CVV isolates. The use of an internal control ensured that possible inefficiency in nucleic acid extraction or PCR inhibition would be detected. |
format | Text |
id | pubmed-2774246 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Elsevier Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-27742462010-10-01 A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() Wang, Heng Nattanmai, Seela Kramer, Laura D. Bernard, Kristen A. Tavakoli, Norma P. Diagn Microbiol Infect Dis Article A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of California (CAL) serogroup viruses and Cache Valley virus (CVV), for use in human surveillance. The targets selected for the assay were the sequences encoding the nucleocapsid protein of CAL and the G1 glycoprotein of CVV. Conserved regions were selected by aligning genetic sequences from various strains available in the GenBank database. Primers and probes were selected in conserved regions. The assay sensitivity was 75 gene copies (gc)/reaction for CAL serogroup viruses and 30 gc/reaction for CVV. The performance of the assay was linear over at least 6 log(10) gc. The assay was specific, given that it did not cross-react with a variety of pathogens. It did, however, detect 11 viruses within the CAL serogroup and 12 CVV isolates. The use of an internal control ensured that possible inefficiency in nucleic acid extraction or PCR inhibition would be detected. Elsevier Inc. 2009-10 2009-09-11 /pmc/articles/PMC2774246/ /pubmed/19748425 http://dx.doi.org/10.1016/j.diagmicrobio.2009.07.001 Text en Copyright © 2009 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Wang, Heng Nattanmai, Seela Kramer, Laura D. Bernard, Kristen A. Tavakoli, Norma P. A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title | A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title_full | A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title_fullStr | A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title_full_unstemmed | A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title_short | A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses() |
title_sort | duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of california serogroup and cache valley viruses() |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774246/ https://www.ncbi.nlm.nih.gov/pubmed/19748425 http://dx.doi.org/10.1016/j.diagmicrobio.2009.07.001 |
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