Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.

Roadkill specimens are an important source of samples for enhanced rabies surveillance (ERS) in areas where other methods of sample collection may not be practical. However, the physical condition of roadkill specimens is unpredictable and, in many circumstances, unsatisfactory for rabies diagnostic...

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Main Authors: Crystal M Gigante, Claire Hartloge, Rene Edgar Condori, Jordona D Kirby, Lauren Hovis, Kathleen M Nelson, Ryan Wallace, Yu Li, Richard B Chipman
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-07-01
Series:PLoS Neglected Tropical Diseases
Online Access:https://doi.org/10.1371/journal.pntd.0013348
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author Crystal M Gigante
Claire Hartloge
Rene Edgar Condori
Jordona D Kirby
Lauren Hovis
Kathleen M Nelson
Ryan Wallace
Yu Li
Richard B Chipman
author_facet Crystal M Gigante
Claire Hartloge
Rene Edgar Condori
Jordona D Kirby
Lauren Hovis
Kathleen M Nelson
Ryan Wallace
Yu Li
Richard B Chipman
author_sort Crystal M Gigante
collection DOAJ
description Roadkill specimens are an important source of samples for enhanced rabies surveillance (ERS) in areas where other methods of sample collection may not be practical. However, the physical condition of roadkill specimens is unpredictable and, in many circumstances, unsatisfactory for rabies diagnostic testing by antigen detection methods. The high sensitivity of real-time reverse transcriptase PCR (RT-PCR) holds promise for rabies diagnostic testing of poor-quality samples. We conducted an evaluation of real-time RT-PCR to detect rabies virus RNA in roadkill samples. A total of 299 specimens were collected from raccoons (n = 232), skunks (n = 46), foxes (n = 17), coyotes (n = 2), a bobcat (n = 1), and a domestic cat (n = 1) across ten states during 2018 - 2021 in the United States. Eight samples (2.7%) were positive using the LN34 pan-lyssavirus real-time RT-PCR assay. These eight rabid animals in areas of high interest for wildlife rabies management would likely not have been identified otherwise. These findings support the use of real-time RT-PCR for samples that would typically be unsuitable for testing by widely used antigenic-based detection methods such as the direct fluorescent antibody test (DFA or FAT) and direct rapid immunohistochemistry test (DRIT).
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1935-2735
language English
publishDate 2025-07-01
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spelling doaj-art-4b0855a53b6d494899e4ab628e6b2a902025-08-20T03:59:35ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352025-07-01197e001334810.1371/journal.pntd.0013348Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.Crystal M GiganteClaire HartlogeRene Edgar CondoriJordona D KirbyLauren HovisKathleen M NelsonRyan WallaceYu LiRichard B ChipmanRoadkill specimens are an important source of samples for enhanced rabies surveillance (ERS) in areas where other methods of sample collection may not be practical. However, the physical condition of roadkill specimens is unpredictable and, in many circumstances, unsatisfactory for rabies diagnostic testing by antigen detection methods. The high sensitivity of real-time reverse transcriptase PCR (RT-PCR) holds promise for rabies diagnostic testing of poor-quality samples. We conducted an evaluation of real-time RT-PCR to detect rabies virus RNA in roadkill samples. A total of 299 specimens were collected from raccoons (n = 232), skunks (n = 46), foxes (n = 17), coyotes (n = 2), a bobcat (n = 1), and a domestic cat (n = 1) across ten states during 2018 - 2021 in the United States. Eight samples (2.7%) were positive using the LN34 pan-lyssavirus real-time RT-PCR assay. These eight rabid animals in areas of high interest for wildlife rabies management would likely not have been identified otherwise. These findings support the use of real-time RT-PCR for samples that would typically be unsuitable for testing by widely used antigenic-based detection methods such as the direct fluorescent antibody test (DFA or FAT) and direct rapid immunohistochemistry test (DRIT).https://doi.org/10.1371/journal.pntd.0013348
spellingShingle Crystal M Gigante
Claire Hartloge
Rene Edgar Condori
Jordona D Kirby
Lauren Hovis
Kathleen M Nelson
Ryan Wallace
Yu Li
Richard B Chipman
Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
PLoS Neglected Tropical Diseases
title Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
title_full Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
title_fullStr Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
title_full_unstemmed Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
title_short Enhanced rabies surveillance in roadkill specimens by real-time RT-PCR.
title_sort enhanced rabies surveillance in roadkill specimens by real time rt pcr
url https://doi.org/10.1371/journal.pntd.0013348
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