Cargando…
Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus
Staphylococcus aureus (S. aureus), including methicillin-resistant S. aureus (MRSA), is one of the most important human pathogens, which is responsible for bacteremia, soft-tissue infections, and food poisoning. Hence, multiple cross displacement amplification (MCDA) is employed to detect all S. aur...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5954800/ https://www.ncbi.nlm.nih.gov/pubmed/29867818 http://dx.doi.org/10.3389/fmicb.2018.00907 |
_version_ | 1783323590230278144 |
---|---|
author | Wang, Yi Yan, Weiqiang Fu, Shanshan Hu, Shoukui Wang, Yan Xu, Jianguo Ye, Changyun |
author_facet | Wang, Yi Yan, Weiqiang Fu, Shanshan Hu, Shoukui Wang, Yan Xu, Jianguo Ye, Changyun |
author_sort | Wang, Yi |
collection | PubMed |
description | Staphylococcus aureus (S. aureus), including methicillin-resistant S. aureus (MRSA), is one of the most important human pathogens, which is responsible for bacteremia, soft-tissue infections, and food poisoning. Hence, multiple cross displacement amplification (MCDA) is employed to detect all S. aureus strains, and differentiates MRSA from methicillin-sensitive S. aureus. Multiplex MCDA (m-MCDA), which targets the nuc gene (S. aureus-specific gene) and mecA gene (encoding penicillin-binding protein-2′), could detect S. aureus strains and identify MRSA within 85 min. Detection of the m-MCDA products is achieved using disposable lateral flow biosensors. A total of 58 strains, including various species of Gram-positive and Gram-negative strains, are used for evaluating and optimizing m-MCDA assays. The optimal amplification condition is found to be 63°C for 40 min, with detection limits at 100 fg DNA/reaction for nuc and mecA genes in the pure cultures, and 10 CFU/tube for nuc and mecA genes in the blood samples. The analytical specificity of m-MCDA assay is of 100%, and no cross-reactions to non-S. aureus strains are produced according to the specificity testing. Particularly, two additional components, including AUDG enzyme and dUTP, are added into the m-MCDA amplification mixtures, which are used for eliminating the unwanted results arising from carryover contamination. Thus, the m-MCDA technique appears to be a simple, rapid, sensitive, and reliable assay to detect all S. aureus strains, and identify MRSA infection for appropriate antibiotic therapy. |
format | Online Article Text |
id | pubmed-5954800 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-59548002018-06-04 Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus Wang, Yi Yan, Weiqiang Fu, Shanshan Hu, Shoukui Wang, Yan Xu, Jianguo Ye, Changyun Front Microbiol Microbiology Staphylococcus aureus (S. aureus), including methicillin-resistant S. aureus (MRSA), is one of the most important human pathogens, which is responsible for bacteremia, soft-tissue infections, and food poisoning. Hence, multiple cross displacement amplification (MCDA) is employed to detect all S. aureus strains, and differentiates MRSA from methicillin-sensitive S. aureus. Multiplex MCDA (m-MCDA), which targets the nuc gene (S. aureus-specific gene) and mecA gene (encoding penicillin-binding protein-2′), could detect S. aureus strains and identify MRSA within 85 min. Detection of the m-MCDA products is achieved using disposable lateral flow biosensors. A total of 58 strains, including various species of Gram-positive and Gram-negative strains, are used for evaluating and optimizing m-MCDA assays. The optimal amplification condition is found to be 63°C for 40 min, with detection limits at 100 fg DNA/reaction for nuc and mecA genes in the pure cultures, and 10 CFU/tube for nuc and mecA genes in the blood samples. The analytical specificity of m-MCDA assay is of 100%, and no cross-reactions to non-S. aureus strains are produced according to the specificity testing. Particularly, two additional components, including AUDG enzyme and dUTP, are added into the m-MCDA amplification mixtures, which are used for eliminating the unwanted results arising from carryover contamination. Thus, the m-MCDA technique appears to be a simple, rapid, sensitive, and reliable assay to detect all S. aureus strains, and identify MRSA infection for appropriate antibiotic therapy. Frontiers Media S.A. 2018-05-09 /pmc/articles/PMC5954800/ /pubmed/29867818 http://dx.doi.org/10.3389/fmicb.2018.00907 Text en Copyright © 2018 Wang, Yan, Fu, Hu, Wang, Xu and Ye. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Wang, Yi Yan, Weiqiang Fu, Shanshan Hu, Shoukui Wang, Yan Xu, Jianguo Ye, Changyun Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title | Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title_full | Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title_fullStr | Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title_full_unstemmed | Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title_short | Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of Staphylococcus aureus and Identification of Methicillin-Resistant S. aureus |
title_sort | multiple cross displacement amplification coupled with nanoparticles-based lateral flow biosensor for detection of staphylococcus aureus and identification of methicillin-resistant s. aureus |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5954800/ https://www.ncbi.nlm.nih.gov/pubmed/29867818 http://dx.doi.org/10.3389/fmicb.2018.00907 |
work_keys_str_mv | AT wangyi multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT yanweiqiang multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT fushanshan multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT hushoukui multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT wangyan multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT xujianguo multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus AT yechangyun multiplecrossdisplacementamplificationcoupledwithnanoparticlesbasedlateralflowbiosensorfordetectionofstaphylococcusaureusandidentificationofmethicillinresistantsaureus |