Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.

Surveillance and detection of polioviruses (PV) remain crucial to monitoring eradication progress. Intratypic differentiation (ITD) using the real-time RT-PCR kit is key to the surveillance workflow, where viruses are screened after cell culture isolation before a subset are verified by sequencing....

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hong Sun, Chelsea Harrington, Nancy Gerloff, Mark Mandelbaum, Stacey Jeffries-Miles, Lea Necitas G Apostol, Ma Anne-Lesley D Valencia, Shahzad Shaukat, Mehar Angez, Deepa K Sharma, Uma P Nalavade, Shailesh D Pawar, Elisabeth Pukuta Simbu, Seta Andriamamonjy, Richter Razafindratsimandresy, Everardo Vega
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/87df276f2f7e4b2e840cccabe7f4dcf8
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:87df276f2f7e4b2e840cccabe7f4dcf8
record_format	dspace
spelling	oai:doaj.org-article:87df276f2f7e4b2e840cccabe7f4dcf82021-12-02T20:18:33ZValidation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.1932-620310.1371/journal.pone.0255795https://doaj.org/article/87df276f2f7e4b2e840cccabe7f4dcf82021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0255795https://doaj.org/toc/1932-6203Surveillance and detection of polioviruses (PV) remain crucial to monitoring eradication progress. Intratypic differentiation (ITD) using the real-time RT-PCR kit is key to the surveillance workflow, where viruses are screened after cell culture isolation before a subset are verified by sequencing. The ITD kit is a series of real-time RT-PCR assays that screens cytopathic effect (CPE)-positive cell cultures using the standard WHO method for virus isolation. Because ITD screening is a critical procedure in the poliovirus identification workflow, validation of performance of real-time PCR platforms is a core requirement for the detection of poliovirus using the ITD kit. In addition, the continual update and improvement of the ITD assays to simplify interpretation in all platforms is necessary to ensure that all real-time machines are capable of detecting positive real-time signals. Four platforms (ABI7500 real-time systems, Bio-Rad CFX96, Stratagene MX3000P, and the Qiagen Rotor-Gene Q) were validated with the ITD kit and a redesigned poliovirus probe. The poliovirus probe in the real-time RT-PCR pan-poliovirus (PanPV) assay was re-designed with a double-quencher (Zen™) to reduce background fluorescence and potential false negatives. The updated PanPV probe was evaluated with a panel consisting of 184 polioviruses and non-polio enteroviruses. To further validate the updated PanPV probe, the new assay was pilot tested in five Global Polio Laboratory Network (GPLN) laboratories (Madagascar, India, Philippines, Pakistan, and Democratic Republic of Congo). The updated PanPV probe performance was shown to reduce background fluorescence and decrease the number of false positives compared to the standard PanPV probe.Hong SunChelsea HarringtonNancy GerloffMark MandelbaumStacey Jeffries-MilesLea Necitas G ApostolMa Anne-Lesley D ValenciaShahzad ShaukatMehar AngezDeepa K SharmaUma P NalavadeShailesh D PawarElisabeth Pukuta SimbuSeta AndriamamonjyRichter RazafindratsimandresyEverardo VegaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 8, p e0255795 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hong Sun Chelsea Harrington Nancy Gerloff Mark Mandelbaum Stacey Jeffries-Miles Lea Necitas G Apostol Ma Anne-Lesley D Valencia Shahzad Shaukat Mehar Angez Deepa K Sharma Uma P Nalavade Shailesh D Pawar Elisabeth Pukuta Simbu Seta Andriamamonjy Richter Razafindratsimandresy Everardo Vega Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
description	Surveillance and detection of polioviruses (PV) remain crucial to monitoring eradication progress. Intratypic differentiation (ITD) using the real-time RT-PCR kit is key to the surveillance workflow, where viruses are screened after cell culture isolation before a subset are verified by sequencing. The ITD kit is a series of real-time RT-PCR assays that screens cytopathic effect (CPE)-positive cell cultures using the standard WHO method for virus isolation. Because ITD screening is a critical procedure in the poliovirus identification workflow, validation of performance of real-time PCR platforms is a core requirement for the detection of poliovirus using the ITD kit. In addition, the continual update and improvement of the ITD assays to simplify interpretation in all platforms is necessary to ensure that all real-time machines are capable of detecting positive real-time signals. Four platforms (ABI7500 real-time systems, Bio-Rad CFX96, Stratagene MX3000P, and the Qiagen Rotor-Gene Q) were validated with the ITD kit and a redesigned poliovirus probe. The poliovirus probe in the real-time RT-PCR pan-poliovirus (PanPV) assay was re-designed with a double-quencher (Zen™) to reduce background fluorescence and potential false negatives. The updated PanPV probe was evaluated with a panel consisting of 184 polioviruses and non-polio enteroviruses. To further validate the updated PanPV probe, the new assay was pilot tested in five Global Polio Laboratory Network (GPLN) laboratories (Madagascar, India, Philippines, Pakistan, and Democratic Republic of Congo). The updated PanPV probe performance was shown to reduce background fluorescence and decrease the number of false positives compared to the standard PanPV probe.
format	article
author	Hong Sun Chelsea Harrington Nancy Gerloff Mark Mandelbaum Stacey Jeffries-Miles Lea Necitas G Apostol Ma Anne-Lesley D Valencia Shahzad Shaukat Mehar Angez Deepa K Sharma Uma P Nalavade Shailesh D Pawar Elisabeth Pukuta Simbu Seta Andriamamonjy Richter Razafindratsimandresy Everardo Vega
author_facet	Hong Sun Chelsea Harrington Nancy Gerloff Mark Mandelbaum Stacey Jeffries-Miles Lea Necitas G Apostol Ma Anne-Lesley D Valencia Shahzad Shaukat Mehar Angez Deepa K Sharma Uma P Nalavade Shailesh D Pawar Elisabeth Pukuta Simbu Seta Andriamamonjy Richter Razafindratsimandresy Everardo Vega
author_sort	Hong Sun
title	Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
title_short	Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
title_full	Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
title_fullStr	Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
title_full_unstemmed	Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.
title_sort	validation of a redesigned pan-poliovirus assay and real-time pcr platforms for the global poliovirus laboratory network.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/87df276f2f7e4b2e840cccabe7f4dcf8
work_keys_str_mv	AT hongsun validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT chelseaharrington validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT nancygerloff validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT markmandelbaum validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT staceyjeffriesmiles validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT leanecitasgapostol validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT maannelesleydvalencia validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT shahzadshaukat validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT meharangez validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT deepaksharma validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT umapnalavade validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT shaileshdpawar validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT elisabethpukutasimbu validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT setaandriamamonjy validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT richterrazafindratsimandresy validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork AT everardovega validationofaredesignedpanpoliovirusassayandrealtimepcrplatformsfortheglobalpolioviruslaboratorynetwork
_version_	1718374294418882560

Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.

Ejemplares similares