Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model.
Genetic analysis of pathogenic organisms is a useful tool for linking human cases together and/or to potential environmental sources. The resulting data can also provide information on evolutionary patterns within a targeted species and phenotypic traits. However, the instruments often used to gener...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2017-12-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC5739503?pdf=render |
| _version_ | 1811219214896201728 |
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| author | Cedar L Mitchell Voahangy Andrianaivoarimanana Rebecca E Colman Joseph Busch Heidie Hornstra-O'Neill Paul S Keim David M Wagner Minoarisoa Rajerison Dawn N Birdsell |
| author_facet | Cedar L Mitchell Voahangy Andrianaivoarimanana Rebecca E Colman Joseph Busch Heidie Hornstra-O'Neill Paul S Keim David M Wagner Minoarisoa Rajerison Dawn N Birdsell |
| author_sort | Cedar L Mitchell |
| collection | DOAJ |
| description | Genetic analysis of pathogenic organisms is a useful tool for linking human cases together and/or to potential environmental sources. The resulting data can also provide information on evolutionary patterns within a targeted species and phenotypic traits. However, the instruments often used to generate genotyping data, such as single nucleotide polymorphisms (SNPs), can be expensive and sometimes require advanced technologies to implement. This places many genotyping tools out of reach for laboratories that do not specialize in genetic studies and/or lack the requisite financial and technological resources. To address this issue, we developed a low cost and low tech genotyping system, termed agarose-MAMA, which combines traditional PCR and agarose gel electrophoresis to target phylogenetically informative SNPs.To demonstrate the utility of this approach for generating genotype data in a resource-constrained area (Madagascar), we designed an agarose-MAMA system targeting previously characterized SNPs within Yersinia pestis, the causative agent of plague. We then used this system to genetically type pathogenic strains of Y. pestis in a Malagasy laboratory not specialized in genetic studies, the Institut Pasteur de Madagascar (IPM). We conducted rigorous assay performance validations to assess potential variation introduced by differing research facilities, reagents, and personnel and found no difference in SNP genotyping results. These agarose-MAMA PCR assays are currently employed as an investigative tool at IPM, providing Malagasy researchers a means to improve the value of their plague epidemiological investigations by linking outbreaks to potential sources through genetic characterization of isolates and to improve understanding of disease ecology that may contribute to a long-term control effort.The success of our study demonstrates that the SNP-based genotyping capacity of laboratories in developing countries can be expanded with manageable financial cost for resource constraint laboratories. This is a practical formula that reduces resource-driven limitations to genetic research and promises to advance global collective knowledge of infectious diseases emanating from resource limited regions of the world. |
| first_indexed | 2024-04-12T07:23:16Z |
| format | Article |
| id | doaj.art-f3e845c6f47141968c29424c333d3a05 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-04-12T07:23:16Z |
| publishDate | 2017-12-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-f3e845c6f47141968c29424c333d3a052022-12-22T03:42:16ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352017-12-011112e000607710.1371/journal.pntd.0006077Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model.Cedar L MitchellVoahangy AndrianaivoarimananaRebecca E ColmanJoseph BuschHeidie Hornstra-O'NeillPaul S KeimDavid M WagnerMinoarisoa RajerisonDawn N BirdsellGenetic analysis of pathogenic organisms is a useful tool for linking human cases together and/or to potential environmental sources. The resulting data can also provide information on evolutionary patterns within a targeted species and phenotypic traits. However, the instruments often used to generate genotyping data, such as single nucleotide polymorphisms (SNPs), can be expensive and sometimes require advanced technologies to implement. This places many genotyping tools out of reach for laboratories that do not specialize in genetic studies and/or lack the requisite financial and technological resources. To address this issue, we developed a low cost and low tech genotyping system, termed agarose-MAMA, which combines traditional PCR and agarose gel electrophoresis to target phylogenetically informative SNPs.To demonstrate the utility of this approach for generating genotype data in a resource-constrained area (Madagascar), we designed an agarose-MAMA system targeting previously characterized SNPs within Yersinia pestis, the causative agent of plague. We then used this system to genetically type pathogenic strains of Y. pestis in a Malagasy laboratory not specialized in genetic studies, the Institut Pasteur de Madagascar (IPM). We conducted rigorous assay performance validations to assess potential variation introduced by differing research facilities, reagents, and personnel and found no difference in SNP genotyping results. These agarose-MAMA PCR assays are currently employed as an investigative tool at IPM, providing Malagasy researchers a means to improve the value of their plague epidemiological investigations by linking outbreaks to potential sources through genetic characterization of isolates and to improve understanding of disease ecology that may contribute to a long-term control effort.The success of our study demonstrates that the SNP-based genotyping capacity of laboratories in developing countries can be expanded with manageable financial cost for resource constraint laboratories. This is a practical formula that reduces resource-driven limitations to genetic research and promises to advance global collective knowledge of infectious diseases emanating from resource limited regions of the world.http://europepmc.org/articles/PMC5739503?pdf=render |
| spellingShingle | Cedar L Mitchell Voahangy Andrianaivoarimanana Rebecca E Colman Joseph Busch Heidie Hornstra-O'Neill Paul S Keim David M Wagner Minoarisoa Rajerison Dawn N Birdsell Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. PLoS Neglected Tropical Diseases |
| title | Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. |
| title_full | Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. |
| title_fullStr | Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. |
| title_full_unstemmed | Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. |
| title_short | Low cost, low tech SNP genotyping tools for resource-limited areas: Plague in Madagascar as a model. |
| title_sort | low cost low tech snp genotyping tools for resource limited areas plague in madagascar as a model |
| url | http://europepmc.org/articles/PMC5739503?pdf=render |
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