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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| Format: | Article |
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmed.2022.831045/full |
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| author | Ju Jiang Ju Jiang Constanza Martínez-Valdebenito Thomas Weitzel Thomas Weitzel Christina M. Farris Gerardo Acosta-Jamett Gerardo Acosta-Jamett Katia Abarca Allen L. Richards Allen L. Richards |
| author_facet | Ju Jiang Ju Jiang Constanza Martínez-Valdebenito Thomas Weitzel Thomas Weitzel Christina M. Farris Gerardo Acosta-Jamett Gerardo Acosta-Jamett Katia Abarca Allen L. Richards Allen L. Richards |
| author_sort | Ju Jiang |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-12T22:30:23Z |
| format | Article |
| id | doaj.art-272d76ccfd1847a29e8a0772302e029f |
| institution | Directory Open Access Journal |
| issn | 2296-858X |
| language | English |
| last_indexed | 2024-12-12T22:30:23Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Medicine |
| spelling | doaj.art-272d76ccfd1847a29e8a0772302e029f2022-12-22T00:09:37ZengFrontiers Media S.A.Frontiers in Medicine2296-858X2022-04-01910.3389/fmed.2022.831045831045Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South AmericaJu Jiang0Ju Jiang1Constanza Martínez-Valdebenito2Thomas Weitzel3Thomas Weitzel4Christina M. Farris5Gerardo Acosta-Jamett6Gerardo Acosta-Jamett7Katia Abarca8Allen L. Richards9Allen L. Richards10Naval Medical Research Center, Silver Spring, MD, United StatesThe Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD, United StatesDepartamento de Enfermedades Infecciosas e Inmunología Pediátrica, Escuela de Medicina, Pontificia Universidad Católica de Chile, Santiago, ChileLaboratorio Clínico, Clínica Alemana de Santiago, Facultad de Medicina Clínica Alemana, Universidad del Desarrollo, Santiago, ChileInstituto de Ciencias e Innovación en Medicina (ICIM), Facultad de Medicina Clínica Alemana, Universidad del Desarrollo, Santiago, ChileNaval Medical Research Center, Silver Spring, MD, United StatesInstituto de Medicina Preventiva Veterinaria, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Valdivia, ChilePrograma de Investigación Aplicada en Fauna Silvestre, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Valdivia, ChileDepartamento de Enfermedades Infecciosas e Inmunología Pediátrica, Escuela de Medicina, Pontificia Universidad Católica de Chile, Santiago, ChileThe Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD, United StatesDepartment of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesScrub 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.https://www.frontiersin.org/articles/10.3389/fmed.2022.831045/fullscrub typhusOrientiaCandidatus Orientia chiloensismolecular diagnostic techniquesquantitative real-time PCR (qPCR)Orien16S |
| spellingShingle | Ju Jiang Ju Jiang Constanza Martínez-Valdebenito Thomas Weitzel Thomas Weitzel Christina M. Farris Gerardo Acosta-Jamett Gerardo Acosta-Jamett Katia Abarca Allen L. Richards Allen L. Richards Development of a New Genus-Specific Quantitative Real-Time PCR Assay for the Diagnosis of Scrub Typhus in South America Frontiers in Medicine scrub typhus Orientia Candidatus Orientia chiloensis molecular diagnostic techniques quantitative real-time PCR (qPCR) Orien16S |
| 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 | scrub typhus Orientia Candidatus Orientia chiloensis molecular diagnostic techniques quantitative real-time PCR (qPCR) Orien16S |
| url | https://www.frontiersin.org/articles/10.3389/fmed.2022.831045/full |
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