RNA Internal Control (IC) for Routine Clinical Diagnostic Real-Time Reverse Transcription-PCR SARS-CoV-2

(RNA Internal Control for Routine rRT-PCR of SARS-CoV-2)

DOI

10.2991/978-94-6463-018-3_12How to use a DOI?

Keywords

RNA; internal control; SARS-CoV-2; Clinical diagnosis; Real time polymerase chain reaction

Abstract

Since the beginning of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that source of a disease (COVID-19) pandemic in Indonesia, laboratories have to applied nucleic acid amplification tests (NAATs), namely real time Reverse-Transcriptase Polymerase Chain Reaction (rRT-PCR) as clinical diagnostic test. This method is extremely high sensitivity and speeds to diagnosis of virus infections. In order to obtain of appropriate in rRT-PCR’s result, internal controls (ICs) has to issued assurance that clinical specimens are successfully amplified and detected. IC is allowing to provides the control being detected only if the target virus is absent and amplification is going well. IC can distinguish extraction failures, rRT-PCR restraint and technical errors relating to each individual sample. IC should be added at the samples prior to extraction. However, some commercial kit for rRT-PCR have available, but their performance for rules to add IC in within RT-PCR procedure has not yet been unassisted evaluated. The objective of this work was to estimated basic analytical of IC preparation for regular diagnostics of COVID-19. We were observed additional IC in RNA extraction and directly into the rRT-PCR reaction. In this study, we were used two commercial kit with conventional RNA extraction method (TIANamp Hi-DNA/RNA Kit) and automated viral RNA isolation (MagMAX™-96 Viral RNA Isolation Kit). The cycle threshold (Ct) were observed in treatment with additional IC in RNA extraction process, both of conventional RNA extraction method and automated RNA extraction method. Conversely, Ct were not observed in additional IC directly in rRT-PCR reaction. We conclude that IC should be added into sample in RNA extraction process.

Copyright

© 2023 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Maharani Pertiwi Koentjoro
AU  - Shamsul Bahari Shamsudin
AU  - Bastiana Bermawi
AU  - Endah Prayekti
AU  - Endry Nugroho Prasetyo
PY  - 2022
DA  - 2022/12/15
TI  - RNA Internal Control (IC) for Routine Clinical Diagnostic Real-Time Reverse Transcription-PCR SARS-CoV-2
BT  - Proceedings of the First International Conference on Medical Technology (ICoMTech 2021)
PB  - Atlantis Press
SP  - 86
EP  - 92
SN  - 2468-5739
UR  - https://doi.org/10.2991/978-94-6463-018-3_12
DO  - 10.2991/978-94-6463-018-3_12
ID  - PertiwiKoentjoro2022
ER  -