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Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts
BACKGROUND: Multiple yeast species can cause human disease, involving superficial to deep‐seated infections. Treatment of these infections depends on the accurate identification of causative agents; however, reliable methods are not available in many laboratories, especially not in resource‐limited...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9102616/ https://www.ncbi.nlm.nih.gov/pubmed/35318737 http://dx.doi.org/10.1002/jcla.24370 |
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author | Aboutalebian, Shima Mahmoudi, Shahram Charsizadeh, Arezoo Nikmanesh, Bahram Hosseini, Mahnaz Mirhendi, Hossein |
author_facet | Aboutalebian, Shima Mahmoudi, Shahram Charsizadeh, Arezoo Nikmanesh, Bahram Hosseini, Mahnaz Mirhendi, Hossein |
author_sort | Aboutalebian, Shima |
collection | PubMed |
description | BACKGROUND: Multiple yeast species can cause human disease, involving superficial to deep‐seated infections. Treatment of these infections depends on the accurate identification of causative agents; however, reliable methods are not available in many laboratories, especially not in resource‐limited settings. Here, a new multiplex assay for rapid and low‐cost identification of pathogenic yeasts is described. METHODS: A two‐step multiplex assay named YEAST PLEX that comprises of four tubes and identifies 17 clinically important common to rare yeasts was designed and evaluated. The set also provides PCR amplicon of unidentified species for direct sequencing. The specificity of YEAST PLEX was tested using 28 reference strains belonging to 17 species and 101 DNA samples of clinically important non‐target bacteria, parasites, and fungi as well as human genomic DNA. The method was further analyzed using 203 previously identified and 89 unknown clinical yeast isolates. Moreover, the method was tested for its ability to identify mixed yeast colonies by using 18 mixed suspensions of two or three species. RESULTS: YEAST PLEX was able to identify all the target species without any non‐specific PCR products. When compared to PCR‐sequencing/MALDI‐TOF, the results of YEAST PLEX were in 100% agreement. Regarding the 89 unknown clinical isolates, random isolates were selected and subjected to PCR‐sequencing. The results of sequencing were in agreement with those of YEAST PLEX. Furthermore, this method was able to correctly identify all yeasts in mixed suspensions. CONCLUSION: YEAST PLEX is an accurate, low‐cost, and rapid method for identification of yeasts, with applicability, especially in developing countries. |
format | Online Article Text |
id | pubmed-9102616 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-91026162022-05-18 Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts Aboutalebian, Shima Mahmoudi, Shahram Charsizadeh, Arezoo Nikmanesh, Bahram Hosseini, Mahnaz Mirhendi, Hossein J Clin Lab Anal Research Articles BACKGROUND: Multiple yeast species can cause human disease, involving superficial to deep‐seated infections. Treatment of these infections depends on the accurate identification of causative agents; however, reliable methods are not available in many laboratories, especially not in resource‐limited settings. Here, a new multiplex assay for rapid and low‐cost identification of pathogenic yeasts is described. METHODS: A two‐step multiplex assay named YEAST PLEX that comprises of four tubes and identifies 17 clinically important common to rare yeasts was designed and evaluated. The set also provides PCR amplicon of unidentified species for direct sequencing. The specificity of YEAST PLEX was tested using 28 reference strains belonging to 17 species and 101 DNA samples of clinically important non‐target bacteria, parasites, and fungi as well as human genomic DNA. The method was further analyzed using 203 previously identified and 89 unknown clinical yeast isolates. Moreover, the method was tested for its ability to identify mixed yeast colonies by using 18 mixed suspensions of two or three species. RESULTS: YEAST PLEX was able to identify all the target species without any non‐specific PCR products. When compared to PCR‐sequencing/MALDI‐TOF, the results of YEAST PLEX were in 100% agreement. Regarding the 89 unknown clinical isolates, random isolates were selected and subjected to PCR‐sequencing. The results of sequencing were in agreement with those of YEAST PLEX. Furthermore, this method was able to correctly identify all yeasts in mixed suspensions. CONCLUSION: YEAST PLEX is an accurate, low‐cost, and rapid method for identification of yeasts, with applicability, especially in developing countries. John Wiley and Sons Inc. 2022-03-22 /pmc/articles/PMC9102616/ /pubmed/35318737 http://dx.doi.org/10.1002/jcla.24370 Text en © 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Aboutalebian, Shima Mahmoudi, Shahram Charsizadeh, Arezoo Nikmanesh, Bahram Hosseini, Mahnaz Mirhendi, Hossein Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title | Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title_full | Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title_fullStr | Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title_full_unstemmed | Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title_short | Multiplex size marker (YEAST PLEX) for rapid and accurate identification of pathogenic yeasts |
title_sort | multiplex size marker (yeast plex) for rapid and accurate identification of pathogenic yeasts |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9102616/ https://www.ncbi.nlm.nih.gov/pubmed/35318737 http://dx.doi.org/10.1002/jcla.24370 |
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