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
Molecular Cancer Biology Group, Zoology Department, Faculty of Science, Ain Shams University, Cairo, 11566, Egypt.
Virology and Immunology Unit, Cancer Biology Department, National Cancer Institute, Cairo University, Cairo, Egypt.
AMB Express. 2022 Jun 9;12(1):70. 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.
全球范围内分子生物学耗材短缺现象激增。这包括过滤吸头、核酸纯化试剂盒、聚合酶、逆转录酶以及病毒诊断试剂盒中包含的各类试剂。在发展中国家，问题更为严峻，因为几乎没有资本企业涉足这类产业。所以，我们的目标是开发一种适用、功能良好、可与商业产品相媲美且价格实惠的内部病毒RNA纯化方案。我们寻找了一些已发表的和商业的RNA纯化方法来建立病毒RNA提取的内部方案。相应地制备了溶液。此外，还对线性化聚丙烯酰胺（LPA）载体进行了评估。将添加了LPA载体的内部溶液的整体设置与QIAamp病毒RNA微型试剂盒溶液进行了比较。我们的结果表明，自制溶液中的线性化聚丙烯酰胺（LPA）载体与大多数商业试剂盒中使用的聚A载体相当。此外，RNA纯化溶液的整体设置确实达到了病毒RNA纯化的目的。使用一名感染新冠病毒患者的痰液也证实了这一结果。我们的实验最终得到了一种价格实惠的自制病毒RNA纯化溶液。
