A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples
Abstract Mosquito-borne diseases pose ongoing global health concerns, demanding more cost-efficient methods to detect pathogens to support enhanced surveillance efforts. This study introduces an adapted TRIzol-based high-throughput RNA extraction protocol, tailored for the detection of California se...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-52534-1 |
| _version_ | 1797274515674234880 |
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| author | Marc Avramov Vanessa Gallo Antonia Gross David R. Lapen Antoinette Ludwig Catherine I. Cullingham |
| author_facet | Marc Avramov Vanessa Gallo Antonia Gross David R. Lapen Antoinette Ludwig Catherine I. Cullingham |
| author_sort | Marc Avramov |
| collection | DOAJ |
| description | Abstract Mosquito-borne diseases pose ongoing global health concerns, demanding more cost-efficient methods to detect pathogens to support enhanced surveillance efforts. This study introduces an adapted TRIzol-based high-throughput RNA extraction protocol, tailored for the detection of California serogroup viruses in pooled mosquito samples in a rapid and cost-effective manner. This approach provided consistent RNA yields and sensitive viral detection relative to two commercial extraction kits (QIAGEN RNeasy Mini Kit and MACHEREY–NAGEL NucleoSpin RNA Plus Kit). The incorporation of a user-friendly and non-spiking-based RT-qPCR internal control designed for the 18S rRNA gene in mosquitoes minimizes potential false positives/negatives, improving the fidelity of viral detection outcomes. Effective RNA yields, purity, and successful target amplification across 25 mosquito species and varied pool sizes (1–50 mosquitoes per tube) affirm the reliability of our approach. The extraction method is cost-effective, with an incurred cost of $0.58 CAD per sample, in contrast to the $5.25 CAD cost per sample of the two kits, rendering it promising for mosquito-borne disease surveillance initiatives. |
| first_indexed | 2024-03-07T15:00:29Z |
| format | Article |
| id | doaj.art-77f40a32c8254eaa86f3d458951c17c8 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-07T15:00:29Z |
| publishDate | 2024-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-77f40a32c8254eaa86f3d458951c17c82024-03-05T19:12:24ZengNature PortfolioScientific Reports2045-23222024-01-011411810.1038/s41598-024-52534-1A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samplesMarc Avramov0Vanessa Gallo1Antonia Gross2David R. Lapen3Antoinette Ludwig4Catherine I. Cullingham5Department of Biology, Carleton UniversityDepartment of Biology, Carleton UniversityDepartment of Biology, Carleton UniversityOttawa Research and Development Centre, Agriculture and Agri-Food CanadaNational Microbiology Laboratory Branch, Public Health Agency of CanadaDepartment of Biology, Carleton UniversityAbstract Mosquito-borne diseases pose ongoing global health concerns, demanding more cost-efficient methods to detect pathogens to support enhanced surveillance efforts. This study introduces an adapted TRIzol-based high-throughput RNA extraction protocol, tailored for the detection of California serogroup viruses in pooled mosquito samples in a rapid and cost-effective manner. This approach provided consistent RNA yields and sensitive viral detection relative to two commercial extraction kits (QIAGEN RNeasy Mini Kit and MACHEREY–NAGEL NucleoSpin RNA Plus Kit). The incorporation of a user-friendly and non-spiking-based RT-qPCR internal control designed for the 18S rRNA gene in mosquitoes minimizes potential false positives/negatives, improving the fidelity of viral detection outcomes. Effective RNA yields, purity, and successful target amplification across 25 mosquito species and varied pool sizes (1–50 mosquitoes per tube) affirm the reliability of our approach. The extraction method is cost-effective, with an incurred cost of $0.58 CAD per sample, in contrast to the $5.25 CAD cost per sample of the two kits, rendering it promising for mosquito-borne disease surveillance initiatives.https://doi.org/10.1038/s41598-024-52534-1 |
| spellingShingle | Marc Avramov Vanessa Gallo Antonia Gross David R. Lapen Antoinette Ludwig Catherine I. Cullingham A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples Scientific Reports |
| title | A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples |
| title_full | A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples |
| title_fullStr | A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples |
| title_full_unstemmed | A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples |
| title_short | A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples |
| title_sort | cost effective rna extraction and rt qpcr approach to detect california serogroup viruses from pooled mosquito samples |
| url | https://doi.org/10.1038/s41598-024-52534-1 |
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