Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae.
<h4>Background</h4>Globally, over 800 000 children under five die each year from infectious diseases caused by Streptococcus pneumoniae. To understand genetic relatedness between isolates, study transmission routes, assess the impact of human interventions e.g. vaccines, and determine in...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0121870 |
| _version_ | 1818837862999654400 |
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| author | Rachael E Rayner John Savill Louise M Hafner Flavia Huygens |
| author_facet | Rachael E Rayner John Savill Louise M Hafner Flavia Huygens |
| author_sort | Rachael E Rayner |
| collection | DOAJ |
| description | <h4>Background</h4>Globally, over 800 000 children under five die each year from infectious diseases caused by Streptococcus pneumoniae. To understand genetic relatedness between isolates, study transmission routes, assess the impact of human interventions e.g. vaccines, and determine infection sources, genotyping methods are required. The 'gold standard' genotyping method, Multi-Locus Sequence Typing (MLST), is useful for long-term and global studies. Another genotyping method, Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA), has emerged as a more discriminatory, inexpensive and faster technique; however there is no universally accepted method and it is currently suitable for short-term and localised epidemiology studies. Currently Australia has no national MLST database, nor has it adopted any MLVA method for short-term or localised studies. This study aims to improve S. pneumoniae genotyping methods by modifying the existing MLVA techniques to be more discriminatory, faster, cheaper and technically less demanding than previously published MLVA methods and MLST.<h4>Methods</h4>Four different MLVA protocols, including a modified method, were applied to 317 isolates of serotyped invasive S. pneumoniae isolated from sterile body sites of Queensland children under 15 years from 2007-2012. MLST was applied to 202 isolates for comparison.<h4>Results</h4>The modified MLVA4 is significantly more discriminatory than the 'gold standard' MLST method. MLVA4 has similar discrimination compared to other MLVA techniques in this study). The failure to amplify particular loci in previous MLVA methods were minimised in MLVA4. Failure to amplify BOX-13 and Spneu19 were found to be serotype specific.<h4>Conclusion</h4>We have modified a highly discriminatory MLVA technique for genotyping Queensland invasive S. pneumoniae. MLVA4 has the ability to enhance our understanding of the pneumococcal epidemiology and the changing genetics of the pneumococcus in localised and short-term studies. |
| first_indexed | 2024-12-19T03:29:15Z |
| format | Article |
| id | doaj.art-68971d5702ee4e0cb3bd1f36393649e2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T03:29:15Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-68971d5702ee4e0cb3bd1f36393649e22022-12-21T20:37:32ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01104e012187010.1371/journal.pone.0121870Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae.Rachael E RaynerJohn SavillLouise M HafnerFlavia Huygens<h4>Background</h4>Globally, over 800 000 children under five die each year from infectious diseases caused by Streptococcus pneumoniae. To understand genetic relatedness between isolates, study transmission routes, assess the impact of human interventions e.g. vaccines, and determine infection sources, genotyping methods are required. The 'gold standard' genotyping method, Multi-Locus Sequence Typing (MLST), is useful for long-term and global studies. Another genotyping method, Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA), has emerged as a more discriminatory, inexpensive and faster technique; however there is no universally accepted method and it is currently suitable for short-term and localised epidemiology studies. Currently Australia has no national MLST database, nor has it adopted any MLVA method for short-term or localised studies. This study aims to improve S. pneumoniae genotyping methods by modifying the existing MLVA techniques to be more discriminatory, faster, cheaper and technically less demanding than previously published MLVA methods and MLST.<h4>Methods</h4>Four different MLVA protocols, including a modified method, were applied to 317 isolates of serotyped invasive S. pneumoniae isolated from sterile body sites of Queensland children under 15 years from 2007-2012. MLST was applied to 202 isolates for comparison.<h4>Results</h4>The modified MLVA4 is significantly more discriminatory than the 'gold standard' MLST method. MLVA4 has similar discrimination compared to other MLVA techniques in this study). The failure to amplify particular loci in previous MLVA methods were minimised in MLVA4. Failure to amplify BOX-13 and Spneu19 were found to be serotype specific.<h4>Conclusion</h4>We have modified a highly discriminatory MLVA technique for genotyping Queensland invasive S. pneumoniae. MLVA4 has the ability to enhance our understanding of the pneumococcal epidemiology and the changing genetics of the pneumococcus in localised and short-term studies.https://doi.org/10.1371/journal.pone.0121870 |
| spellingShingle | Rachael E Rayner John Savill Louise M Hafner Flavia Huygens Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. PLoS ONE |
| title | Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. |
| title_full | Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. |
| title_fullStr | Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. |
| title_full_unstemmed | Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. |
| title_short | Modified MLVA for Genotyping Queensland Invasive Streptococcus pneumoniae. |
| title_sort | modified mlva for genotyping queensland invasive streptococcus pneumoniae |
| url | https://doi.org/10.1371/journal.pone.0121870 |
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