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Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens

During a large variety of common pathogens, E. coli, P. aeruginosa, MRSA, MRCNS, V. parahaemolyticus, L. monocytogenes and Salmonella are the leading pathogens responsible for large number of human infections and diseases. In this study, a high flux screening based on nucleic acid isothermal amplifi...

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Autores principales: Zhong, Huamin, Deng, Hongwei, Li, Ming, Zhong, Huahong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096746/
https://www.ncbi.nlm.nih.gov/pubmed/32862325
http://dx.doi.org/10.1007/s00449-020-02423-4
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author Zhong, Huamin
Deng, Hongwei
Li, Ming
Zhong, Huahong
author_facet Zhong, Huamin
Deng, Hongwei
Li, Ming
Zhong, Huahong
author_sort Zhong, Huamin
collection PubMed
description During a large variety of common pathogens, E. coli, P. aeruginosa, MRSA, MRCNS, V. parahaemolyticus, L. monocytogenes and Salmonella are the leading pathogens responsible for large number of human infections and diseases. In this study, a high flux screening based on nucleic acid isothermal amplification technique has been developed. For the 8 common pathogens, species-specific targets had been selected and analyzed for their unique specificity. After optimization, separate LAMP reaction assays had been bioprocessed and integrated into one systematic detection platform, including 8 strips (PCR tubes) and 96-well plates. Eight standard strains verified for the accuracy. Application of the established high flux screening platform was used for detection for 48 samples in 4 different 96-well plates, with 2 groups of 2 operators using double-blind procedure. The accuracy of 100% was obtained, with the total time consumption as 66–75 min (for 12 samples detection on 8 different pathogens). As concluded, through the bioprocess of the systematic platform based on LAMP technique, it’s been demonstrated to be capable of simultaneous detection of 8 pathogens, with high sensitivity, specificity, rapidity and convenience.
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spelling pubmed-80967462021-05-05 Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens Zhong, Huamin Deng, Hongwei Li, Ming Zhong, Huahong Bioprocess Biosyst Eng Research Paper During a large variety of common pathogens, E. coli, P. aeruginosa, MRSA, MRCNS, V. parahaemolyticus, L. monocytogenes and Salmonella are the leading pathogens responsible for large number of human infections and diseases. In this study, a high flux screening based on nucleic acid isothermal amplification technique has been developed. For the 8 common pathogens, species-specific targets had been selected and analyzed for their unique specificity. After optimization, separate LAMP reaction assays had been bioprocessed and integrated into one systematic detection platform, including 8 strips (PCR tubes) and 96-well plates. Eight standard strains verified for the accuracy. Application of the established high flux screening platform was used for detection for 48 samples in 4 different 96-well plates, with 2 groups of 2 operators using double-blind procedure. The accuracy of 100% was obtained, with the total time consumption as 66–75 min (for 12 samples detection on 8 different pathogens). As concluded, through the bioprocess of the systematic platform based on LAMP technique, it’s been demonstrated to be capable of simultaneous detection of 8 pathogens, with high sensitivity, specificity, rapidity and convenience. Springer Berlin Heidelberg 2020-08-29 2021 /pmc/articles/PMC8096746/ /pubmed/32862325 http://dx.doi.org/10.1007/s00449-020-02423-4 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Paper
Zhong, Huamin
Deng, Hongwei
Li, Ming
Zhong, Huahong
Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title_full Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title_fullStr Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title_full_unstemmed Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title_short Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
title_sort bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096746/
https://www.ncbi.nlm.nih.gov/pubmed/32862325
http://dx.doi.org/10.1007/s00449-020-02423-4
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