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Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS

Candida tropicalis is one of the few Candida species besides Candida albicans that is able to produce true hyphae. At present, the commonly used clinical methods for the identification of this organism are traditional fungal culture, CTB staining, and color development. Polymerase chain reaction (PC...

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Autores principales: Wang, Lei, Xu, Aiguo, Zhou, Ping, Zhao, Mengdi, Xu, Chenglai, Wang, Yan, Wang, Kun, Wang, Fang, Miao, Yongchang, Zhao, Weiguo, Gao, Xuzhu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9301490/
https://www.ncbi.nlm.nih.gov/pubmed/35873165
http://dx.doi.org/10.3389/fcimb.2022.898186
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author Wang, Lei
Xu, Aiguo
Zhou, Ping
Zhao, Mengdi
Xu, Chenglai
Wang, Yan
Wang, Kun
Wang, Fang
Miao, Yongchang
Zhao, Weiguo
Gao, Xuzhu
author_facet Wang, Lei
Xu, Aiguo
Zhou, Ping
Zhao, Mengdi
Xu, Chenglai
Wang, Yan
Wang, Kun
Wang, Fang
Miao, Yongchang
Zhao, Weiguo
Gao, Xuzhu
author_sort Wang, Lei
collection PubMed
description Candida tropicalis is one of the few Candida species besides Candida albicans that is able to produce true hyphae. At present, the commonly used clinical methods for the identification of this organism are traditional fungal culture, CTB staining, and color development. Polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR) are also used to identify this fungus. Since the course of C. tropicalis infection progresses rapidly, there is an urgent need for rapid, sensitive, real-time field assays to meet the needs of clinical diagnosis. Recombinase polymerase amplification (RPA) combined with lateral flow strip (LFS) can rapidly amplify and visualize target genes within 20 min, and by pre-processing samples from different sources, the entire process can be controlled within 30 min. In this study, RPA-LFS was used to amplify the internal transcribed spacer-2 (ITS2) gene of C. tropicalis, and primer-probe design was optimized by introducing base mismatches to obtain a specific and sensitive primer-probe combination for clinical sample detection. LFS assay for 37 common clinical pathogens was performed, sensitivity and specificity of the detection system was determined, reaction temperature and time were optimized, and 191 actual clinical samples collected from different sources were tested to evaluate the detection performance of the established RPA-LFS system to provide a reliable molecular diagnostic method for the detection of C. tropicalis, the results show that the RPA-LFS system can specifically detect C. tropicalis without cross-reacting with other fungi or bacterial, with a sensitivity of 9.94 CFU/µL, without interference from genomic DNA of other species, at an optimal reaction temperature of 39°C, and the whole reaction process can be controlled within 20 min, and to meet the clinical need for rapid, sensitive, real-time, and portable field testing.
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spelling pubmed-93014902022-07-22 Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS Wang, Lei Xu, Aiguo Zhou, Ping Zhao, Mengdi Xu, Chenglai Wang, Yan Wang, Kun Wang, Fang Miao, Yongchang Zhao, Weiguo Gao, Xuzhu Front Cell Infect Microbiol Cellular and Infection Microbiology Candida tropicalis is one of the few Candida species besides Candida albicans that is able to produce true hyphae. At present, the commonly used clinical methods for the identification of this organism are traditional fungal culture, CTB staining, and color development. Polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR) are also used to identify this fungus. Since the course of C. tropicalis infection progresses rapidly, there is an urgent need for rapid, sensitive, real-time field assays to meet the needs of clinical diagnosis. Recombinase polymerase amplification (RPA) combined with lateral flow strip (LFS) can rapidly amplify and visualize target genes within 20 min, and by pre-processing samples from different sources, the entire process can be controlled within 30 min. In this study, RPA-LFS was used to amplify the internal transcribed spacer-2 (ITS2) gene of C. tropicalis, and primer-probe design was optimized by introducing base mismatches to obtain a specific and sensitive primer-probe combination for clinical sample detection. LFS assay for 37 common clinical pathogens was performed, sensitivity and specificity of the detection system was determined, reaction temperature and time were optimized, and 191 actual clinical samples collected from different sources were tested to evaluate the detection performance of the established RPA-LFS system to provide a reliable molecular diagnostic method for the detection of C. tropicalis, the results show that the RPA-LFS system can specifically detect C. tropicalis without cross-reacting with other fungi or bacterial, with a sensitivity of 9.94 CFU/µL, without interference from genomic DNA of other species, at an optimal reaction temperature of 39°C, and the whole reaction process can be controlled within 20 min, and to meet the clinical need for rapid, sensitive, real-time, and portable field testing. Frontiers Media S.A. 2022-07-07 /pmc/articles/PMC9301490/ /pubmed/35873165 http://dx.doi.org/10.3389/fcimb.2022.898186 Text en Copyright © 2022 Wang, Xu, Zhou, Zhao, Xu, Wang, Wang, Wang, Miao, Zhao and Gao 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 Cellular and Infection Microbiology
Wang, Lei
Xu, Aiguo
Zhou, Ping
Zhao, Mengdi
Xu, Chenglai
Wang, Yan
Wang, Kun
Wang, Fang
Miao, Yongchang
Zhao, Weiguo
Gao, Xuzhu
Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title_full Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title_fullStr Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title_full_unstemmed Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title_short Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS
title_sort rapid detection of candida tropicalis in clinical samples from different sources using rpa-lfs
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9301490/
https://www.ncbi.nlm.nih.gov/pubmed/35873165
http://dx.doi.org/10.3389/fcimb.2022.898186
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