High-quality RT-PCR with chemically modified RNA controls.

Abstract

In detecting infectious diseases, such as coronavirus 2019 (COVID-19), real-time reverse-transcription polymerase chain reaction (RT-PCR) is one of the most important technologies for RNA detection and disease diagnosis. To achieve high quality assurance, appropriate positive and negative controls are critical for disease detection using RT-PCR kits. In this study, we have found that commercial kits often adopt DNAs instead of RNAs as the positive controls, which can't report the kit problems in reverse transcription, thereby increasing risk of the false negative results when testing patient samples. To face the challenge, we have proposed and developed the chemically modified RNAs, such as phosphoroselenaote and phosphorothioate RNAs (Se-RNA and S-RNA), as the controls. We have found that while demonstrating the high thermostability, biostability, chemostability and exclusivity (or specificity), both Se-RNA and S-RNA can be fine templates for reverse transcription, indicating their potentials as both positive and negative controls for RT-PCR kits.

• Authors: Guangcheng Luo, Jun Zhang, Shun Zhang, Bei Hu, Lillian Hu, Zhen Huang
• Journal: Talanta (Talanta) Vol. 224 Pg. 121850 (Mar 01 2021) ISSN: 1873-3573 [Electronic] Netherlands
• PMID: 33379066 (Publication Type: Journal Article)
• Copyright: Copyright © 2020. Published by Elsevier B.V.

Chemical References

• DNA, Viral
• RNA, Viral
• Reagent Kits, Diagnostic

Topics

• COVID-19 (diagnosis)
• COVID-19 Nucleic Acid Testing (instrumentation, methods)
• DNA, Viral (analysis)
• False Negative Reactions
• Humans
• RNA Stability
• RNA, Viral (analysis, chemistry)
• Reagent Kits, Diagnostic
• Real-Time Polymerase Chain Reaction (instrumentation, methods)
• Reverse Transcriptase Polymerase Chain Reaction (instrumentation, methods)
• SARS-CoV-2 (genetics)