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Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples

This study evaluated the performance of the Maxwell 16 System (Promega) for extraction of influenza virus (flu-v) RNA from diverse samples compared to a classical manual method (QIAamp Kit, QIAGEN). Following extraction by the two methods, all samples were analyzed by Real-time RT-PCR. Results revea...

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Detalles Bibliográficos
Autores principales: Liu, Hongbo, Gan, Yan, Yang, Bo, Weng, Hui, Huang, Chunmei, Yang, Daofeng, Lei, Ping, Shen, Guanxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3483271/
https://www.ncbi.nlm.nih.gov/pubmed/23144730
http://dx.doi.org/10.1371/journal.pone.0048094
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author Liu, Hongbo
Gan, Yan
Yang, Bo
Weng, Hui
Huang, Chunmei
Yang, Daofeng
Lei, Ping
Shen, Guanxin
author_facet Liu, Hongbo
Gan, Yan
Yang, Bo
Weng, Hui
Huang, Chunmei
Yang, Daofeng
Lei, Ping
Shen, Guanxin
author_sort Liu, Hongbo
collection PubMed
description This study evaluated the performance of the Maxwell 16 System (Promega) for extraction of influenza virus (flu-v) RNA from diverse samples compared to a classical manual method (QIAamp Kit, QIAGEN). Following extraction by the two methods, all samples were analyzed by Real-time RT-PCR. Results revealed that the use of the standard Maxwell 16 protocol (Maxwell 16-S) resulted in good linearity and precision across a wide concentration range and higher sensitivity of detection from flu-v stock suspensions than the manual method. Compared with the latter method, Maxwell 16-S extracted RNA more efficiently (higher RNA yield and/or fewer PCR inhibitors) from throat swabs and bronchoalveolar lavage fluids, while both methods performed comparably on fecal samples from human and poultry in terms of overall threshold cycle values and detection rates although the Maxwell 16-S co-purified more inhibitors from fecal samples. The capacity of this system to remove inhibitors from fecal matrix was improved by using a modified Maxwell 16 protocol with a reduced sample input, which eliminated all false-negatives produced by the Maxwell 16-S. These findings suggest that the Maxwell 16 System is suitable for RNA extraction from multiple-source samples for diagnosis of influenza and viral load determination and that a proper reduction in starting sample volume may improve the detection of flu-v from complex matrices such as feces. Additionally, this system allows flexible sample throughput and labor-saving sample processing with little or no risk of cross-contamination.
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spelling pubmed-34832712012-11-09 Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples Liu, Hongbo Gan, Yan Yang, Bo Weng, Hui Huang, Chunmei Yang, Daofeng Lei, Ping Shen, Guanxin PLoS One Research Article This study evaluated the performance of the Maxwell 16 System (Promega) for extraction of influenza virus (flu-v) RNA from diverse samples compared to a classical manual method (QIAamp Kit, QIAGEN). Following extraction by the two methods, all samples were analyzed by Real-time RT-PCR. Results revealed that the use of the standard Maxwell 16 protocol (Maxwell 16-S) resulted in good linearity and precision across a wide concentration range and higher sensitivity of detection from flu-v stock suspensions than the manual method. Compared with the latter method, Maxwell 16-S extracted RNA more efficiently (higher RNA yield and/or fewer PCR inhibitors) from throat swabs and bronchoalveolar lavage fluids, while both methods performed comparably on fecal samples from human and poultry in terms of overall threshold cycle values and detection rates although the Maxwell 16-S co-purified more inhibitors from fecal samples. The capacity of this system to remove inhibitors from fecal matrix was improved by using a modified Maxwell 16 protocol with a reduced sample input, which eliminated all false-negatives produced by the Maxwell 16-S. These findings suggest that the Maxwell 16 System is suitable for RNA extraction from multiple-source samples for diagnosis of influenza and viral load determination and that a proper reduction in starting sample volume may improve the detection of flu-v from complex matrices such as feces. Additionally, this system allows flexible sample throughput and labor-saving sample processing with little or no risk of cross-contamination. Public Library of Science 2012-10-29 /pmc/articles/PMC3483271/ /pubmed/23144730 http://dx.doi.org/10.1371/journal.pone.0048094 Text en © 2012 Liu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Liu, Hongbo
Gan, Yan
Yang, Bo
Weng, Hui
Huang, Chunmei
Yang, Daofeng
Lei, Ping
Shen, Guanxin
Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title_full Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title_fullStr Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title_full_unstemmed Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title_short Performance Evaluation of the Maxwell 16 System for Extraction of Influenza Virus RNA from Diverse Samples
title_sort performance evaluation of the maxwell 16 system for extraction of influenza virus rna from diverse samples
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3483271/
https://www.ncbi.nlm.nih.gov/pubmed/23144730
http://dx.doi.org/10.1371/journal.pone.0048094
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