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Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica

Yersinia enterocolitica remains a threat to public health, and a sensitive detection method is a prerequisite due to its complicated diagnosis associated with slow growth. Recently, aptamer-based detection systems have played a vital role in the development of simple, rapid, sensitive, and specific...

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Autores principales: Shoaib, Muhammad, Shehzad, Aamir, Mukama, Omar, Raza, Husnain, Niazi, Sobia, Khan, Imran Mahmood, Ali, Barkat, Akhtar, Wasim, Wang, Zhouping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9055141/
https://www.ncbi.nlm.nih.gov/pubmed/35516186
http://dx.doi.org/10.1039/d0ra00683a
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author Shoaib, Muhammad
Shehzad, Aamir
Mukama, Omar
Raza, Husnain
Niazi, Sobia
Khan, Imran Mahmood
Ali, Barkat
Akhtar, Wasim
Wang, Zhouping
author_facet Shoaib, Muhammad
Shehzad, Aamir
Mukama, Omar
Raza, Husnain
Niazi, Sobia
Khan, Imran Mahmood
Ali, Barkat
Akhtar, Wasim
Wang, Zhouping
author_sort Shoaib, Muhammad
collection PubMed
description Yersinia enterocolitica remains a threat to public health, and a sensitive detection method is a prerequisite due to its complicated diagnosis associated with slow growth. Recently, aptamer-based detection systems have played a vital role in the development of simple, rapid, sensitive, and specific detection methods. Herein, highly specific ssDNA aptamers were screened against Y. enterocolitica at the different growth stages by whole cell-SELEX. Cells at different growth stages were harvested and incubated with an ssDNA library to get an enriched pool of specific aptamer candidates. After the 10(th) round of SELEX, the enriched pool was sequenced and grouped into seven families based on homology and similarity of the secondary structure. Flow cytometry analysis revealed that the aptamers M1, M5, and M7 with K(d) values of 37.93 ± 7.88 nM, 74.96 ± 21.34 nM, and 73.02 ± 18.76 nM had the highest affinity and specificity to the target, respectively. The selected aptamers showed binding to the different growth stages of Y. enterocolitica with a significant increase in the gated fluorescence. Our aptamer selection strategy is convenient, and the developed aptamer can be useful for an accurate and reliable detection system.
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spelling pubmed-90551412022-05-04 Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica Shoaib, Muhammad Shehzad, Aamir Mukama, Omar Raza, Husnain Niazi, Sobia Khan, Imran Mahmood Ali, Barkat Akhtar, Wasim Wang, Zhouping RSC Adv Chemistry Yersinia enterocolitica remains a threat to public health, and a sensitive detection method is a prerequisite due to its complicated diagnosis associated with slow growth. Recently, aptamer-based detection systems have played a vital role in the development of simple, rapid, sensitive, and specific detection methods. Herein, highly specific ssDNA aptamers were screened against Y. enterocolitica at the different growth stages by whole cell-SELEX. Cells at different growth stages were harvested and incubated with an ssDNA library to get an enriched pool of specific aptamer candidates. After the 10(th) round of SELEX, the enriched pool was sequenced and grouped into seven families based on homology and similarity of the secondary structure. Flow cytometry analysis revealed that the aptamers M1, M5, and M7 with K(d) values of 37.93 ± 7.88 nM, 74.96 ± 21.34 nM, and 73.02 ± 18.76 nM had the highest affinity and specificity to the target, respectively. The selected aptamers showed binding to the different growth stages of Y. enterocolitica with a significant increase in the gated fluorescence. Our aptamer selection strategy is convenient, and the developed aptamer can be useful for an accurate and reliable detection system. The Royal Society of Chemistry 2020-06-30 /pmc/articles/PMC9055141/ /pubmed/35516186 http://dx.doi.org/10.1039/d0ra00683a Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Shoaib, Muhammad
Shehzad, Aamir
Mukama, Omar
Raza, Husnain
Niazi, Sobia
Khan, Imran Mahmood
Ali, Barkat
Akhtar, Wasim
Wang, Zhouping
Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title_full Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title_fullStr Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title_full_unstemmed Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title_short Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica
title_sort selection of potential aptamers for specific growth stage detection of yersinia enterocolitica
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9055141/
https://www.ncbi.nlm.nih.gov/pubmed/35516186
http://dx.doi.org/10.1039/d0ra00683a
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