Development of a Novel Viral RNA Extraction Method

Abstract

Limitations of commercial RNA extraction kits make them inappropriate for use in a clinical setting. This project develops and evaluates a point-of-care process for simple and rapid extraction of RNA from clinical samples. The novel RNA extraction method does not require laboratory equipment and is simpler and faster. In this thesis the proposed method is compared to three existing commercial extraction kits. Comparisons were made using RT-PCR of extraction solutions obtained after adding a known number of copies of respiratory syncytial virus (RSV) RNA to buffer or negative patient samples and using previously stored RSV positive patient samples. A comparison of viral RNA extraction from buffer found that our initial design of the proposed method achieved extraction efficiency of 80 to 90% of commercial kits. Extraction performance using previously stored RSV positive patient samples ranged from ~15 to 67% of commercial kits. With further optimization this approach would likely have a significant impact on moving genome-based detection methods into a clinical setting.