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High throughput MLVA-16 typing for Brucella based on the microfluidics technology
BACKGROUND: Brucellosis, a zoonosis caused by the genus Brucella, has been eradicated in Northern Europe, Australia, the USA and Canada, but remains endemic in most areas of the world. The strain and biovar typing of Brucella field samples isolated in outbreaks is useful for tracing back source of i...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3078841/ https://www.ncbi.nlm.nih.gov/pubmed/21435217 http://dx.doi.org/10.1186/1471-2180-11-60 |
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author | De Santis, Riccardo Ciammaruconi, Andrea Faggioni, Giovanni Fillo, Silvia Gentile, Bernardina Di Giannatale, Elisabetta Ancora, Massimo Lista, Florigio |
author_facet | De Santis, Riccardo Ciammaruconi, Andrea Faggioni, Giovanni Fillo, Silvia Gentile, Bernardina Di Giannatale, Elisabetta Ancora, Massimo Lista, Florigio |
author_sort | De Santis, Riccardo |
collection | PubMed |
description | BACKGROUND: Brucellosis, a zoonosis caused by the genus Brucella, has been eradicated in Northern Europe, Australia, the USA and Canada, but remains endemic in most areas of the world. The strain and biovar typing of Brucella field samples isolated in outbreaks is useful for tracing back source of infection and may be crucial for discriminating naturally occurring outbreaks versus bioterrorist events, being Brucella a potential biological warfare agent. In the last years MLVA-16 has been described for Brucella spp. genotyping. The MLVA band profiles may be resolved by different techniques i.e. the manual agarose gels, the capillary electrophoresis sequencing systems or the microfluidic Lab-on-Chip electrophoresis. In this paper we described a high throughput system of MLVA-16 typing for Brucella spp. by using of the microfluidics technology. RESULTS: The Caliper LabChip 90 equipment was evaluated for MLVA-16 typing of sixty-three Brucella samples. Furthermore, in order to validate the system, DNA samples previously resolved by sequencing system and Agilent technology, were de novo genotyped. The comparison of the MLVA typing data obtained by the Caliper equipment and those previously obtained by the other analysis methods showed a good correlation. However the outputs were not accurate as the Caliper DNA fragment sizes showed discrepancies compared with real data and a conversion table from observed to expected data was created. CONCLUSION: In this paper we described the MLVA-16 using a rapid, sophisticated microfluidics technology for detection of amplification product sizes. The comparison of the MLVA typing data produced by Caliper LabChip 90 system with the data obtained by different techniques showed a general concordance of the results. Furthermore this platform represents a significant improvement in terms of handling, data acquiring, computational efficiency and rapidity, allowing to perform the strain genotyping in a time equal to one sixth respect to other microfluidics systems as e.g. the Agilent 2100 bioanalyzer. Finally, this platform can be considered a valid alternative to standard genotyping techniques, particularly useful dealing with a large number of samples in short time. These data confirmed that this technology represents a significative advancement in high-throughput accurate Brucella genotyping. |
format | Text |
id | pubmed-3078841 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-30788412011-04-19 High throughput MLVA-16 typing for Brucella based on the microfluidics technology De Santis, Riccardo Ciammaruconi, Andrea Faggioni, Giovanni Fillo, Silvia Gentile, Bernardina Di Giannatale, Elisabetta Ancora, Massimo Lista, Florigio BMC Microbiol Methodology Article BACKGROUND: Brucellosis, a zoonosis caused by the genus Brucella, has been eradicated in Northern Europe, Australia, the USA and Canada, but remains endemic in most areas of the world. The strain and biovar typing of Brucella field samples isolated in outbreaks is useful for tracing back source of infection and may be crucial for discriminating naturally occurring outbreaks versus bioterrorist events, being Brucella a potential biological warfare agent. In the last years MLVA-16 has been described for Brucella spp. genotyping. The MLVA band profiles may be resolved by different techniques i.e. the manual agarose gels, the capillary electrophoresis sequencing systems or the microfluidic Lab-on-Chip electrophoresis. In this paper we described a high throughput system of MLVA-16 typing for Brucella spp. by using of the microfluidics technology. RESULTS: The Caliper LabChip 90 equipment was evaluated for MLVA-16 typing of sixty-three Brucella samples. Furthermore, in order to validate the system, DNA samples previously resolved by sequencing system and Agilent technology, were de novo genotyped. The comparison of the MLVA typing data obtained by the Caliper equipment and those previously obtained by the other analysis methods showed a good correlation. However the outputs were not accurate as the Caliper DNA fragment sizes showed discrepancies compared with real data and a conversion table from observed to expected data was created. CONCLUSION: In this paper we described the MLVA-16 using a rapid, sophisticated microfluidics technology for detection of amplification product sizes. The comparison of the MLVA typing data produced by Caliper LabChip 90 system with the data obtained by different techniques showed a general concordance of the results. Furthermore this platform represents a significant improvement in terms of handling, data acquiring, computational efficiency and rapidity, allowing to perform the strain genotyping in a time equal to one sixth respect to other microfluidics systems as e.g. the Agilent 2100 bioanalyzer. Finally, this platform can be considered a valid alternative to standard genotyping techniques, particularly useful dealing with a large number of samples in short time. These data confirmed that this technology represents a significative advancement in high-throughput accurate Brucella genotyping. BioMed Central 2011-03-24 /pmc/articles/PMC3078841/ /pubmed/21435217 http://dx.doi.org/10.1186/1471-2180-11-60 Text en Copyright ©2011 De Santis et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Article De Santis, Riccardo Ciammaruconi, Andrea Faggioni, Giovanni Fillo, Silvia Gentile, Bernardina Di Giannatale, Elisabetta Ancora, Massimo Lista, Florigio High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title | High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title_full | High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title_fullStr | High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title_full_unstemmed | High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title_short | High throughput MLVA-16 typing for Brucella based on the microfluidics technology |
title_sort | high throughput mlva-16 typing for brucella based on the microfluidics technology |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3078841/ https://www.ncbi.nlm.nih.gov/pubmed/21435217 http://dx.doi.org/10.1186/1471-2180-11-60 |
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