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Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America

Scrub typhus is a potentially severe rickettsiosis, caused by Orientia tsutsugamushi in the Asia-Pacific region. Recently, however, two distinct pathogens, “Candidatus Orientia chuto” and “Candidatus Orientia chiloensis”, have been discovered in the Middle East and South America, respectively. Since...

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Autores principales: Jiang, Ju, Martínez-Valdebenito, Constanza, Weitzel, Thomas, Farris, Christina M., Acosta-Jamett, Gerardo, Abarca, Katia, Richards, Allen L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9095740/
https://www.ncbi.nlm.nih.gov/pubmed/35573006
http://dx.doi.org/10.3389/fmed.2022.831045
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author Jiang, Ju
Martínez-Valdebenito, Constanza
Weitzel, Thomas
Farris, Christina M.
Acosta-Jamett, Gerardo
Abarca, Katia
Richards, Allen L.
author_facet Jiang, Ju
Martínez-Valdebenito, Constanza
Weitzel, Thomas
Farris, Christina M.
Acosta-Jamett, Gerardo
Abarca, Katia
Richards, Allen L.
author_sort Jiang, Ju
collection PubMed
description Scrub typhus is a potentially severe rickettsiosis, caused by Orientia tsutsugamushi in the Asia-Pacific region. Recently, however, two distinct pathogens, “Candidatus Orientia chuto” and “Candidatus Orientia chiloensis”, have been discovered in the Middle East and South America, respectively. Since the novel pathogens differ significantly from O. tsutsugamushi, many established diagnostic methods are unreliable. This work describes the development and validation of a new quantitative real-time PCR (qPCR) assay (Orien16S) for the detection of all known Orientia species. Based on a 94 bp sequence of the 16S rRNA gene (rrs), Orien16S recognized DNA samples from O. tsutsugamushi (n = 41), Ca. O. chiloensis (n = 5), and Ca. O. chuto (n = 1), but was negative for DNA preparations from closely related rickettsiae and other members of the order Rickettsiales (n = 22) as well as unrelated bacterial species (n = 11). After its implementation in Chile, the assay was verified, correctly identifying all tested eschar and buffy coat samples (n = 28) of clinical suspected cases. Furthermore, Orien16S detected Orientia DNA in trombiculid mites collected in endemic regions in southern Chile. The presented novel qPCR assay provides a useful tool for detecting Orientia and diagnosing scrub typhus from all geographical regions.
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spelling pubmed-90957402022-05-13 Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America Jiang, Ju Martínez-Valdebenito, Constanza Weitzel, Thomas Farris, Christina M. Acosta-Jamett, Gerardo Abarca, Katia Richards, Allen L. Front Med (Lausanne) Medicine Scrub typhus is a potentially severe rickettsiosis, caused by Orientia tsutsugamushi in the Asia-Pacific region. Recently, however, two distinct pathogens, “Candidatus Orientia chuto” and “Candidatus Orientia chiloensis”, have been discovered in the Middle East and South America, respectively. Since the novel pathogens differ significantly from O. tsutsugamushi, many established diagnostic methods are unreliable. This work describes the development and validation of a new quantitative real-time PCR (qPCR) assay (Orien16S) for the detection of all known Orientia species. Based on a 94 bp sequence of the 16S rRNA gene (rrs), Orien16S recognized DNA samples from O. tsutsugamushi (n = 41), Ca. O. chiloensis (n = 5), and Ca. O. chuto (n = 1), but was negative for DNA preparations from closely related rickettsiae and other members of the order Rickettsiales (n = 22) as well as unrelated bacterial species (n = 11). After its implementation in Chile, the assay was verified, correctly identifying all tested eschar and buffy coat samples (n = 28) of clinical suspected cases. Furthermore, Orien16S detected Orientia DNA in trombiculid mites collected in endemic regions in southern Chile. The presented novel qPCR assay provides a useful tool for detecting Orientia and diagnosing scrub typhus from all geographical regions. Frontiers Media S.A. 2022-04-27 /pmc/articles/PMC9095740/ /pubmed/35573006 http://dx.doi.org/10.3389/fmed.2022.831045 Text en Copyright © 2022 Jiang, Martínez-Valdebenito, Weitzel, Farris, Acosta-Jamett, Abarca and Richards. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Medicine
Jiang, Ju
Martínez-Valdebenito, Constanza
Weitzel, Thomas
Farris, Christina M.
Acosta-Jamett, Gerardo
Abarca, Katia
Richards, Allen L.
Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title_full Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title_fullStr Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title_full_unstemmed Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title_short Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America
title_sort development of a new genus-specific quantitative real-time pcr assay for the diagnosis of scrub typhus in south america
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9095740/
https://www.ncbi.nlm.nih.gov/pubmed/35573006
http://dx.doi.org/10.3389/fmed.2022.831045
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