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Virology

Diagnostic Assay Development for Poliovirus Eradication

Nancy Gerloff, Hong Sun, Mark Mandelbaum, Chelsea Maher, W. Allan Nix, Sohail Zaidi, Shahzad Shaukat, Lerato Seakamela, Uma P. Nalavade, Deepa K. Sharma, M. Steven Oberste, Everardo Vega
Angela M. Caliendo, Editor
Nancy Gerloff
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Hong Sun
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Mark Mandelbaum
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Chelsea Maher
bIHRC, Inc., Atlanta, Georgia, USA
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W. Allan Nix
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Sohail Zaidi
cNational Institute of Health, Islamabad, Pakistan
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Shahzad Shaukat
cNational Institute of Health, Islamabad, Pakistan
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Lerato Seakamela
dNational Institute for Communicable Diseases, Johannesburg, South Africa
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Uma P. Nalavade
eEnterovirus Research Centre, Mumbai, India
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Deepa K. Sharma
eEnterovirus Research Centre, Mumbai, India
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M. Steven Oberste
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Everardo Vega
aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
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Angela M. Caliendo
Rhode Island Hospital
Roles: Editor
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DOI: 10.1128/JCM.01624-17
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ABSTRACT

With poliovirus eradication nearing, few pockets of active wild poliovirus (WPV) transmission remain in the world. Intratypic differentiation (ITD) plays a crucial part in laboratory surveillance as the molecular detection method that can identify and distinguish wild and vaccine-like polioviruses isolated from acute flaccid paralysis cases or environmental sources. The need to detect new variants of WPV serotype 1 (WPV1) and the containment of all serotype 2 polioviruses (PV2) in 2015 required changes to the previous version of the method. The ITD version 5.0 is a set of six real-time reverse transcription-PCR (rRT-PCR) assays that serve as accurate diagnostic tools to easily detect and differentiate PV serotypes and genotypes. We describe the creation and properties of quantitation standards, including 16 control RNA transcripts and nine plaque-isolated viruses. All ITD rRT-PCR assays were validated using these standards, and the limits of detection were determined for each assay. We designed and pilot tested two new assays targeting recently circulating WPV1 genotypes and all PV2 viruses. The WPV1 assay had 99.1% specificity and 100% sensitivity, and the PV2 assay had 97.7% specificity and 92% sensitivity. Before proceeding to the next step in the global poliovirus eradication program, we needed to gain a better understanding of the performance of the ITD 5.0 suite of molecular assays and their limits of detection and specificities. The findings and conclusions in this evaluation serve as building blocks for future development work.

FOOTNOTES

    • Received 12 October 2017.
    • Returned for modification 3 November 2017.
    • Accepted 1 December 2017.
    • Accepted manuscript posted online 6 December 2017.
  • Supplemental material for this article may be found at https://doi.org/10.1128/JCM.01624-17.

  • Copyright © 2018 American Society for Microbiology.

All Rights Reserved.

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Diagnostic Assay Development for Poliovirus Eradication
Nancy Gerloff, Hong Sun, Mark Mandelbaum, Chelsea Maher, W. Allan Nix, Sohail Zaidi, Shahzad Shaukat, Lerato Seakamela, Uma P. Nalavade, Deepa K. Sharma, M. Steven Oberste, Everardo Vega
Journal of Clinical Microbiology Jan 2018, 56 (2) e01624-17; DOI: 10.1128/JCM.01624-17

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Diagnostic Assay Development for Poliovirus Eradication
Nancy Gerloff, Hong Sun, Mark Mandelbaum, Chelsea Maher, W. Allan Nix, Sohail Zaidi, Shahzad Shaukat, Lerato Seakamela, Uma P. Nalavade, Deepa K. Sharma, M. Steven Oberste, Everardo Vega
Journal of Clinical Microbiology Jan 2018, 56 (2) e01624-17; DOI: 10.1128/JCM.01624-17
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KEYWORDS

diagnostic assay
molecular typing
polio eradication
poliovirus
rRT-PCR
real-time reverse transcription-PCR
serotype identification

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