Lee Alex H F, Gessert Steven F, Chen Yutao, Sergeev Nikolay V, Haghiri Babak
Theranos Inc., 7333 Gateway Blvd, Newark, CA 94560, USA.
Heliyon. 2018 Mar 14;4(3):e00572. doi: 10.1016/j.heliyon.2018.e00572. eCollection 2018 Mar.
In this work, a robust synthetic pathway for magnetic core preparation and silica surface coating of magnetic microparticles is presented. Silica-coated magnetic particles are widely used to extract DNA and RNA from various biological samples. We present a novel route for the synthesis of iron oxide silica particles (FeO@Silica) and demonstrate their performance for extracting ZIKA viral RNA from serum. The iron (II, III) oxide (FeO), magnetite core is first prepared by ammonia neutralization of ferrous and ferric chloride aqueous solution under argon, followed by the addition of citrate salt to stabilize the surface of the resultant magnetic nanospheres. After this one-pot, two-step synthesis, the magnetic nanospheres are consumed during silica coating by hydrolysis of tetraethoxysilane (TEOS) under alkaline condition. The final product is a sphere-like magnetic aggregate with a size range of 1-2 micron. By simply suspending the magnetic aggregates in guanidinium chloride solution, the silica surface can be prepared for RNA binding. The RNA extraction efficiency was evaluated by extracting ZIKA viral RNA from serum followed by a PCR-based assay. The data indicate excellent recovery of target RNA and removal of PCR inhibitors. This manufacturing procedure for the silica coated microparticles provides a low-cost, effective and ready for scale-up method whose performance is equivalent to commercial alternatives such as magnetic silica surface particles for DNA and RNA sample preparations. The cost of the clinical assays could be largely decreased due to the 100 fold reduction in cost by replacing the commercially available magnetic particles with the developed material for RNA extraction.
在这项工作中,我们展示了一种用于制备磁性微粒子磁芯并进行二氧化硅表面包覆的稳健合成途径。二氧化硅包覆的磁性颗粒被广泛用于从各种生物样品中提取DNA和RNA。我们提出了一种合成氧化铁二氧化硅颗粒(FeO@二氧化硅)的新方法,并展示了它们从血清中提取寨卡病毒RNA的性能。首先通过在氩气氛围下用氨水中和氯化亚铁和氯化铁水溶液来制备氧化亚铁(FeO)磁芯,然后添加柠檬酸盐以稳定所得磁性纳米球的表面。在这个一锅两步合成之后,在碱性条件下通过四乙氧基硅烷(TEOS)的水解,磁性纳米球在二氧化硅包覆过程中被消耗。最终产物是一种尺寸范围为1 - 2微米的球状磁性聚集体。通过简单地将磁性聚集体悬浮在氯化铵溶液中,就可以使二氧化硅表面用于RNA结合。通过从血清中提取寨卡病毒RNA然后进行基于PCR的检测来评估RNA提取效率。数据表明目标RNA回收率高且PCR抑制剂被去除。这种二氧化硅包覆微粒子的制造方法提供了一种低成本、有效且易于扩大规模的方法,其性能与用于DNA和RNA样品制备的商业替代品如磁性二氧化硅表面颗粒相当。由于用所开发的用于RNA提取的材料替代市售磁性颗粒可使成本降低100倍,临床检测成本可能会大幅降低。