Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards
FTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/23/12915 |
| _version_ | 1797507684375724032 |
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| author | Ahmed Elnagar Timm C. Harder Sandra Blome Martin Beer Bernd Hoffmann |
| author_facet | Ahmed Elnagar Timm C. Harder Sandra Blome Martin Beer Bernd Hoffmann |
| author_sort | Ahmed Elnagar |
| collection | DOAJ |
| description | FTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the validation, dilution series of <i>African swine fever virus</i> (ASFV) positive EDTA blood and <i>Influenza A virus</i> (IAV) positive allantoic fluid were used. Based on our data, we conclude that direct PCR amplification without the need for additional nucleic acid extraction and purification could be suitable and more convenient for ASFV DNA release from FTA cards. In contrast, IAV RNA loads can be amplified from FTA card punches if a standard extraction procedure including a lysis step is applied. These differences between the amplifiable viral DNA and RNA after releasing and extraction are not influenced by the type of commercial FTA card or the eleven different nucleic acid releasing procedures used for the comparative analyses. In general, different commercial FTA cards were successfully used for the storage and recovery of the ASFV and IAV genetic material suitable for PCR. Nevertheless, the usage of optimized nucleic acid releasing protocols could improve the recovery of the viral genome of both viruses. Here, the application of Chelex<sup>®</sup> Resin 100 buffer mixed with 1 × Tris EDTA buffer (TE, pH 8.0) or with TED 10 (TE buffer and Dimethylsulfoxid) delivered the best results and can be used as a universal method for releasing viral DNA and RNA from FTA cards. |
| first_indexed | 2024-03-10T04:52:56Z |
| format | Article |
| id | doaj.art-0ee761f29eb5414182592ddcb8d54316 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:52:56Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0ee761f29eb5414182592ddcb8d543162023-11-23T02:30:12ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-11-0122231291510.3390/ijms222312915Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA CardsAhmed Elnagar0Timm C. Harder1Sandra Blome2Martin Beer3Bernd Hoffmann4Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald, GermanyFTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the validation, dilution series of <i>African swine fever virus</i> (ASFV) positive EDTA blood and <i>Influenza A virus</i> (IAV) positive allantoic fluid were used. Based on our data, we conclude that direct PCR amplification without the need for additional nucleic acid extraction and purification could be suitable and more convenient for ASFV DNA release from FTA cards. In contrast, IAV RNA loads can be amplified from FTA card punches if a standard extraction procedure including a lysis step is applied. These differences between the amplifiable viral DNA and RNA after releasing and extraction are not influenced by the type of commercial FTA card or the eleven different nucleic acid releasing procedures used for the comparative analyses. In general, different commercial FTA cards were successfully used for the storage and recovery of the ASFV and IAV genetic material suitable for PCR. Nevertheless, the usage of optimized nucleic acid releasing protocols could improve the recovery of the viral genome of both viruses. Here, the application of Chelex<sup>®</sup> Resin 100 buffer mixed with 1 × Tris EDTA buffer (TE, pH 8.0) or with TED 10 (TE buffer and Dimethylsulfoxid) delivered the best results and can be used as a universal method for releasing viral DNA and RNA from FTA cards.https://www.mdpi.com/1422-0067/22/23/12915<i>African swine fever virus</i><i>Influenza A virus</i>nucleic acid releaseDNA/RNA isolationdirect PCR amplificationFTA cards |
| spellingShingle | Ahmed Elnagar Timm C. Harder Sandra Blome Martin Beer Bernd Hoffmann Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards International Journal of Molecular Sciences <i>African swine fever virus</i> <i>Influenza A virus</i> nucleic acid release DNA/RNA isolation direct PCR amplification FTA cards |
| title | Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards |
| title_full | Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards |
| title_fullStr | Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards |
| title_full_unstemmed | Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards |
| title_short | Optimizing Release of Nucleic Acids of <i>African Swine Fever Virus</i> and <i>Influenza A Virus</i> from FTA Cards |
| title_sort | optimizing release of nucleic acids of i african swine fever virus i and i influenza a virus i from fta cards |
| topic | <i>African swine fever virus</i> <i>Influenza A virus</i> nucleic acid release DNA/RNA isolation direct PCR amplification FTA cards |
| url | https://www.mdpi.com/1422-0067/22/23/12915 |
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