Hongjaisee S., Jabjainai Y., Sakset S., Preechasuth K., Ngo-Giang-Huong Nicole, Khamduang W. (2022). Comparison of simple RNA extraction methods for molecular diagnosis of hepatitis C virus in plasma. Diagnostics, 12 (7), p. 1599 [6 p.].
Titre du document
Comparison of simple RNA extraction methods for molecular diagnosis of hepatitis C virus in plasma
Hongjaisee S., Jabjainai Y., Sakset S., Preechasuth K., Ngo-Giang-Huong Nicole, Khamduang W.
Source
Diagnostics, 2022,
12 (7), p. 1599 [6 p.]
Nucleic acid extraction from biological samples is an important step for hepatitis C virus (HCV) diagnosis. However, such extractions are mostly based on silica-based column methodologies, which may limit their application for on-site diagnosis. A simple, rapid, and field-deployable method for RNA extraction is still needed. In this study, we evaluated the efficacy of four simple RNA extraction methods for the detection of HCV in plasma samples: a silica-membrane-based method, a magnetic-beads-based method, boiling with diethyl pyrocarbonate (DEPC)-treated distilled water, and using a commercial lysis buffer. HCV RNA was detected using both real-time reverse transcription polymerase chain reaction (RT-PCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP). Using real-time RT-PCR, extracted RNA from the silica-membrane-based and magnetic-beads-based methods had a 100% detection rate for RNA extraction from plasma. Using RT-LAMP, extracted RNA from the silica-membrane-based method showed a 66% detection rate, while the magnetic-beads-based method had a 62% detection rate. In summary, magnetic-beads-based extraction can be used as an alternative RNA extraction method for on-site HCV detection. Boiling with DEPC-treated distilled water was not appropriate for low HCV load samples, and boiling with a lysis buffer was not recommended.
