Comparison of Simple RNA Extraction Methods for Molecular Diagnosis of Hepatitis C Virus in Plasma

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 e...

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Bibliographic Details
Published in:Diagnostics (Basel) 2022-06, Vol.12 (7), p.1599
Main Authors: Hongjaisee, Sayamon, Jabjainai, Yosita, Sakset, Suthasinee, Preechasuth, Kanya, Ngo-Giang-Huong, Nicole, Khamduang, Woottichai
Format: Article
Language:English
Subjects:
Acids
Communication
Diagnosis
HCV
Health aspects
Hepatitis C
Identification and classification
Methods
molecular diagnosis
Molecular diagnostic techniques
Nucleic acids
Plasma
Polymerase chain reaction
RNA
RNA extraction
RT-LAMP
RT-PCR
Separation
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Summary: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.
ISSN:2075-4418
2075-4418
DOI:10.3390/diagnostics12071599