Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples
| Autor: | Christopher E. Jensen, Anders Fomsgaard, Maiken Worsøe Rosenstierne |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Lysis Clinical samples General Chemical Engineering General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Blood Specimen Collection/methods Issue 136 Nucleic Acids Humans Tube (fluid conveyance) Nucleic Acid Amplification Techniques/methods Diagnostics Whole blood Immunology and Infection Blood Specimen Collection Chromatography General Immunology and Microbiology Elution Chemistry Nucleic acid extraction General Neuroscience Extraction (chemistry) Virus 030104 developmental biology Bedside Nucleic Acids/isolation & purification Nucleic acid Blood Collection Tube Nucleic Acid Amplification Techniques Blood sampling |
| Zdroj: | J Vis Exp Rosenstierne, M W, Jensen, C E & Fomsgaard, A 2018, ' Rapid, safe, and simple manual bedside nucleic acid extraction for the detection of virus in whole blood samples ', Journal of Visualized Experiments, vol. 2018, no. 136, e58001 . https://doi.org/10.3791/58001 |
| ISSN: | 1940-087X |
| Popis: | The rapid diagnosis of an infection is essential for the outbreak management, risk containment, and patient care. We have previously shown a method for the rapid bedside inactivation of the Ebola virus during blood sampling for safe nucleic acid (NA) tests by adding a commercial lysis/ binding buffer directly into the vacuum blood collection tubes. Using this bedside inactivation approach, we have developed a safe, rapid, and simplified bedside NA extraction method for the subsequent detection of a virus in lysis/binding buffer-inactivated whole blood. The NA extraction is directly performed in the blood collection tubes and requires no equipment or electricity. After the blood is collected into the lysis/binding buffer, the contents are mixed by flipping the tube by hand, and the mixture is incubated for 20 min at room temperature. Magnetic glass particles (MGPs) are added to the tube, and the contents are mixed by flipping the collection tube by hand. The MGPs are then collected on the side of the blood collection tube using a magnetic holder or a magnet and a rubber band. The MGPs are washed three times, and after the addition of elution buffer directly into the collection tube, the NAs are ready for NA tests, such as qPCR or isothermal loop amplification (LAMP), without the removal of the MGPs from the reaction. The NA extraction method is not dependent on any laboratory facilities and can easily be used anywhere (e.g., in field hospitals and hospital isolation wards). When this NA extraction method is combined with LAMP and a portable instrument, a diagnosis can be obtained within 40 min of the blood collection. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|