RNA提取
色谱法
溶解
核糖核酸
化学
检出限
萃取(化学)
逆转录聚合酶链式反应
逆转录酶
丙型肝炎病毒
裂解缓冲液
膜
核酸
蒸馏水
病毒
分子生物学
病毒学
生物
生物化学
信使核糖核酸
基因
作者
Sayamon Hongjaisee,Yosita Jabjainai,Suthasinee Sakset,Kanya Preechasuth,Nicole Ngo‐Giang‐Huong,Woottichai Khamduang
出处
期刊:Diagnostics
[Multidisciplinary Digital Publishing Institute]
日期:2022-06-30
卷期号:12 (7): 1599-1599
被引量:7
标识
DOI:10.3390/diagnostics12071599
摘要
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.
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