In-house protocol: spin-based viral RNA purification

• Published in: AMB Express Vol. 12; no. 1; p. 70
• Main Authors: Abdelfattah, Mahmoud M., Osman, Ahmed M., Elnagar, Mohamed A., Ibrahim, Mohamed F., Albert, Magdy, M.Talal, Maya, Abdel Fattah, Nasra F., Loutfy, Samah A., Helwa, Reham
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 09.06.2022; Springer Nature B.V; SpringerOpen
• Subjects: Acids; Biomedical and Life Sciences; Biotechnology; Developing countries; Ethanol; Experiments; In-house protocol; LDCs; Life Sciences; Linearization; LPA carrier; Measles; Microbial Genetics and Genomics; Microbiology; Molecular biology; Nucleic acids; Original; Original Article; Polyacrylamide; Purification; Reagents; Ribonucleic acid; RNA; RNA purification; Severe acute respiratory syndrome coronavirus 2; Sputum; Vero cell line; Viral RNA; Vero cell line; Viral RNA; LPA carrier; In-house protocol; RNA purification
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/s13568-022-01413-x

A worldwide shortage of molecular biology consumables is in surge. This includes filter tips, nucleic acid purification kits, polymerases, reverse-transcriptase, and different types of reagents which are included in viral diagnostic kits. In developing countries, the problem is even worse, since there is few capital enterprise to adopt this kind of industry. So, our aim is to develop a suitable, functional, comparable to commercial ones, and affordable in-house protocol to purify viral RNA. We sought some published and commercial RNA purification solutions to set-up an in-house protocol for viral RNA extraction. Solution was prepared accordingly. Also, LPA (linearized polyacrylamide) carrier was evaluated. The whole setting of in-house solutions with addition of LPA carrier was compared to QIAamp viral RNA minikit solutions. Our results showed that linearized polyacrylamide (LPA) carrier in homemade solutions is comparable to poly A carrier which is used in the most commercial kit. In addition, the whole setting of RNA purification solutions did achieve the purpose of viral RNA purification. Also, the result was confirmed using sputum of a Sars-Cov2 infected patient. Our experiments did end up with an affordable homemade solutions for viral RNA purification.