A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses

A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of St. Louis encephalitis virus (SLEV) and eastern equine encephalitis virus (EEEV), for use in human and vector surveillance. The respective targets selected for the assay were the...

Descripción completa

Detalles Bibliográficos
Autores principales: Hull, Rene, Nattanmai, Seela, Kramer, Laura D., Bernard, Kristen A., Tavakoli, Norma P.
Formato: Texto
Lenguaje:English
Publicado: Elsevier Biomedical 2008
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2615585/
https://www.ncbi.nlm.nih.gov/pubmed/18715737
http://dx.doi.org/10.1016/j.diagmicrobio.2008.07.004
_version_ 1782163340145983488
author Hull, Rene
Nattanmai, Seela
Kramer, Laura D.
Bernard, Kristen A.
Tavakoli, Norma P.
author_facet Hull, Rene
Nattanmai, Seela
Kramer, Laura D.
Bernard, Kristen A.
Tavakoli, Norma P.
author_sort Hull, Rene
collection PubMed
description A duplex TaqMan real-time reverse transcriptase polymerase chain reaction (PCR) assay was developed for the detection of St. Louis encephalitis virus (SLEV) and eastern equine encephalitis virus (EEEV), for use in human and vector surveillance. The respective targets selected for the assay were the conserved NS5 and E1 genes of the 2 viruses. Because of the insufficient number of NS5 sequences from SLEV strains in the GenBank database, we determined the sequence of an approximately 1-kb region for each of 25 strains of SLEV to select primers and probes in a conserved region. Our assay has a sensitivity of 5 gene copies (gc)/reaction for EEEV and 10 gc/reaction for SLEV, and its performance is linear for at least 6 log(10) gc. The assay is specific and detected all strains of SLEV (69) and EEEV (12) that were tested. An internal control ensures detection of efficient nucleic acid extraction and possible PCR inhibition.
format Text
id pubmed-2615585
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher Elsevier Biomedical
record_format MEDLINE/PubMed
spelling pubmed-26155852009-11-01 A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses Hull, Rene 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 St. Louis encephalitis virus (SLEV) and eastern equine encephalitis virus (EEEV), for use in human and vector surveillance. The respective targets selected for the assay were the conserved NS5 and E1 genes of the 2 viruses. Because of the insufficient number of NS5 sequences from SLEV strains in the GenBank database, we determined the sequence of an approximately 1-kb region for each of 25 strains of SLEV to select primers and probes in a conserved region. Our assay has a sensitivity of 5 gene copies (gc)/reaction for EEEV and 10 gc/reaction for SLEV, and its performance is linear for at least 6 log(10) gc. The assay is specific and detected all strains of SLEV (69) and EEEV (12) that were tested. An internal control ensures detection of efficient nucleic acid extraction and possible PCR inhibition. Elsevier Biomedical 2008-11 2008-08-20 /pmc/articles/PMC2615585/ /pubmed/18715737 http://dx.doi.org/10.1016/j.diagmicrobio.2008.07.004 Text en 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
Hull, Rene
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 St. Louis encephalitis and eastern equine encephalitis viruses
title A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses
title_full A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses
title_fullStr A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses
title_full_unstemmed A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses
title_short A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of St. Louis encephalitis and eastern equine encephalitis viruses
title_sort duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of st. louis encephalitis and eastern equine encephalitis viruses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2615585/
https://www.ncbi.nlm.nih.gov/pubmed/18715737
http://dx.doi.org/10.1016/j.diagmicrobio.2008.07.004
work_keys_str_mv AT hullrene aduplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT nattanmaiseela aduplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT kramerlaurad aduplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT bernardkristena aduplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT tavakolinormap aduplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT hullrene duplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT nattanmaiseela duplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT kramerlaurad duplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT bernardkristena duplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses
AT tavakolinormap duplexrealtimereversetranscriptasepolymerasechainreactionassayforthedetectionofstlouisencephalitisandeasternequineencephalitisviruses

Ejemplares similares