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Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis
Candida parapsilosis is a common cause of candidiasis among hospitalized patients, often surpassing Candida albicans. Due to the recent increase in C. parapsilosis infections, there is an urgent need for rapid, sensitive, and real-time on-site detection of nucleic acids for timely diagnosis of candi...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10195124/ https://www.ncbi.nlm.nih.gov/pubmed/37202545 http://dx.doi.org/10.1007/s00284-023-03318-9 |
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author | Zhu, Bo Wang, Lei Lu, Yingzhi Chen, Cheng Wang, Kun Zhang, Lei |
author_facet | Zhu, Bo Wang, Lei Lu, Yingzhi Chen, Cheng Wang, Kun Zhang, Lei |
author_sort | Zhu, Bo |
collection | PubMed |
description | Candida parapsilosis is a common cause of candidiasis among hospitalized patients, often surpassing Candida albicans. Due to the recent increase in C. parapsilosis infections, there is an urgent need for rapid, sensitive, and real-time on-site detection of nucleic acids for timely diagnosis of candidiasis. We developed an assay for detection of C. parapsilosis by combining recombinase polymerase amplification (RPA) with a lateral flow strip (LFS). The RPA-LFS assay was used to amplify the beta-1,3-glucan synthase catalytic subunit 2 (FKS2) gene of C. parapsilosis with a primer–probe set optimized by introducing base mismatches (four bases modified by the probe and one by the reverse primer) to achieve specific and sensitive detection of clinical samples. The RPA assays can rapidly amplify and visualize a target gene within 30 min, while the entire process can be completed within 40 min by pre-processing the sample. The product of RPA has two chemical labels, FITC and Biotin, of the amplification product can be carefully on the strip. The sensitivity and specificity of the RPA-LFS assay were determined by analysis of 35 common clinical pathogens and 281 clinical samples against quantitative PCR. The results confirmed that the proposed RPA-LFS assay is a reliable molecular diagnostic method for the detection of C. parapsilosis to meet the urgent need for rapid, specific, sensitive, and portable field testing. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00284-023-03318-9. |
format | Online Article Text |
id | pubmed-10195124 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-101951242023-05-19 Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis Zhu, Bo Wang, Lei Lu, Yingzhi Chen, Cheng Wang, Kun Zhang, Lei Curr Microbiol Article Candida parapsilosis is a common cause of candidiasis among hospitalized patients, often surpassing Candida albicans. Due to the recent increase in C. parapsilosis infections, there is an urgent need for rapid, sensitive, and real-time on-site detection of nucleic acids for timely diagnosis of candidiasis. We developed an assay for detection of C. parapsilosis by combining recombinase polymerase amplification (RPA) with a lateral flow strip (LFS). The RPA-LFS assay was used to amplify the beta-1,3-glucan synthase catalytic subunit 2 (FKS2) gene of C. parapsilosis with a primer–probe set optimized by introducing base mismatches (four bases modified by the probe and one by the reverse primer) to achieve specific and sensitive detection of clinical samples. The RPA assays can rapidly amplify and visualize a target gene within 30 min, while the entire process can be completed within 40 min by pre-processing the sample. The product of RPA has two chemical labels, FITC and Biotin, of the amplification product can be carefully on the strip. The sensitivity and specificity of the RPA-LFS assay were determined by analysis of 35 common clinical pathogens and 281 clinical samples against quantitative PCR. The results confirmed that the proposed RPA-LFS assay is a reliable molecular diagnostic method for the detection of C. parapsilosis to meet the urgent need for rapid, specific, sensitive, and portable field testing. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00284-023-03318-9. Springer US 2023-05-18 2023 /pmc/articles/PMC10195124/ /pubmed/37202545 http://dx.doi.org/10.1007/s00284-023-03318-9 Text en © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Article Zhu, Bo Wang, Lei Lu, Yingzhi Chen, Cheng Wang, Kun Zhang, Lei Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title | Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title_full | Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title_fullStr | Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title_full_unstemmed | Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title_short | Recombinase Polymerase Amplification Assay with Lateral Flow Strips for Rapid Detection of Candidiasis Due to Candida parapsilosis |
title_sort | recombinase polymerase amplification assay with lateral flow strips for rapid detection of candidiasis due to candida parapsilosis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10195124/ https://www.ncbi.nlm.nih.gov/pubmed/37202545 http://dx.doi.org/10.1007/s00284-023-03318-9 |
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