Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
Here, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, blaCTXM1-like, blaCMY-2, qnrA and qnrS) from e...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016121002818 |
| _version_ | 1818968927723585536 |
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| author | Roberta Tolosi Lisa Carraro Andrea Laconi Alessandra Piccirillo |
| author_facet | Roberta Tolosi Lisa Carraro Andrea Laconi Alessandra Piccirillo |
| author_sort | Roberta Tolosi |
| collection | DOAJ |
| description | Here, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, blaCTXM1-like, blaCMY-2, qnrA and qnrS) from environmental samples (i.e. soil and manure). These five protocols accurately detected and quantified the aforementioned ARGs in complex environmental matrices and represent useful tools for both diagnostic and monitoring activities of resistant bacteria and ARGs into the environment. |
| first_indexed | 2024-12-20T14:12:28Z |
| format | Article |
| id | doaj.art-32e038ebac1a4d21969939b1e99934a6 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-20T14:12:28Z |
| publishDate | 2021-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-32e038ebac1a4d21969939b1e99934a62022-12-21T19:38:07ZengElsevierMethodsX2215-01612021-01-018101488Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samplesRoberta Tolosi0Lisa Carraro1Andrea Laconi2Alessandra Piccirillo3Department of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università 16, Legnaro, Padua 35020, ItalyDepartment of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università 16, Legnaro, Padua 35020, ItalyCorresponding author.; Department of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università 16, Legnaro, Padua 35020, ItalyDepartment of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università 16, Legnaro, Padua 35020, ItalyHere, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, blaCTXM1-like, blaCMY-2, qnrA and qnrS) from environmental samples (i.e. soil and manure). These five protocols accurately detected and quantified the aforementioned ARGs in complex environmental matrices and represent useful tools for both diagnostic and monitoring activities of resistant bacteria and ARGs into the environment.http://www.sciencedirect.com/science/article/pii/S2215016121002818Antimicrobial resistance genesqPCRSYBRGreen |
| spellingShingle | Roberta Tolosi Lisa Carraro Andrea Laconi Alessandra Piccirillo Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples MethodsX Antimicrobial resistance genes qPCR SYBRGreen |
| title | Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| title_full | Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| title_fullStr | Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| title_full_unstemmed | Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| title_short | Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| title_sort | optimization of five qpcr protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples |
| topic | Antimicrobial resistance genes qPCR SYBRGreen |
| url | http://www.sciencedirect.com/science/article/pii/S2215016121002818 |
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