rRT-PCR for SARS-CoV-2: Analytical considerations

Rahbari, Rezgar; Moradi, Nariman; Abdi, Mohammad

doi:10.1016/j.cca.2021.01.011

Cited by 71 publications

(53 citation statements)

References 53 publications

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“…Moreover, in a recent systematic review, it was suggested that up to 54% of COVID-19 patients may have an initial negative RT-PCR result [ 5 ]. There are several potential sources causing these differences, which include sampling, storage and processing, RNA extraction, cDNA synthesis and amplification, as well as post-analytical steps (interpretation and analysis) [ 14 ]. Although similar issues may affect the final result of MS-based assays, our work highlights the advantage of MassARRAY ® technology for SARS-CoV-2 diagnosis ( Table 4 ).…”

Section: Discussionmentioning

confidence: 99%

Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Rybicka

Miłosz

Bielawski

2021

Viruses

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At present, the RT-PCR test remains the gold standard for early diagnosis of SARS-CoV-2. Nevertheless, there is growing evidence demonstrating that this technique may generate false-negative results. Here, we aimed to compare the new mass spectrometry-based assay MassARRAY® SARS-CoV-2 Panel with the RT-PCR diagnostic test approved for clinical use. The study group consisted of 168 suspected patients with symptoms of a respiratory infection. After simultaneous analysis by RT-PCR and mass spectrometry methods, we obtained discordant results for 17 samples (10.12%). Within fifteen samples officially reported as presumptive positive, 13 were positive according to the MS-based assay. Moreover, four samples reported by the officially approved RT-PCR as negative were positive in at least one MS assay. We have successfully demonstrated superior sensitivity of the MS-based assay in SARS-CoV-2 detection, showing that MALDI-TOF MS seems to be ideal for the detection as well as discrimination of mutations within the viral genome.

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Section: Discussionmentioning

confidence: 99%

Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Rybicka

Miłosz

Bielawski

2021

Viruses

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“…This study focuses on applying various RNA extraction methods and RT-qPCR kits to detect SARS-CoV-2. There are three main ways of viral RNA extraction used in diagnostic laboratories worldwide [ 14 ]. The first is entirely automatic and requires special equipment, which is not available in each laboratory.…”

Section: Discussionmentioning

confidence: 99%

“…The primary source of errors leading to improper diagnostic of SARS-CoV-2 was described by Rahbari et al [ 14 ] and divided into three groups: pre-analytical, including sampling steps; analytical, including nucleic acid extraction methods and RT-qPCR assays; and post-analytical, focusing on the misinterpretation of the results. Therefore, it is crucial to choose appropriate research methods and interpret the results.…”

Section: Introductionmentioning

confidence: 99%

Impact of the Nucleic Acid Extraction Method and the RT-qPCR Assay on SARS-CoV-2 Detection in Low-Viral Samples

et al. 2021

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COVID-19 was initially reported in China at the end of 2019 and soon thereafter, in March 2020, the WHO declared it a pandemic. Until October 2021, over 240 million COVID-19 cases were recorded, with 4.9 mln deaths. In order to stop the spread of this disease, it is crucial to monitor and detect any infected person. The etiologic agent of COVID-19 is a novel coronavirus called SARS-CoV-2. The gold standard for the detection of the virus is the RT-qPCR method. This study evaluated two RNA extraction methods and four commercial RT-qPCR assays routinely used in diagnostic laboratories for detecting SARS-CoV-2 in human specimens from the upper respiratory tract. We analyzed a panel of 70 clinical samples with varying RNA loads. Our study demonstrated the significant impact of the diagnostic methods selected by the laboratory on the SARS-CoV-2 detection in clinical specimens with low viral loads.

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“…The variants were neutralized with convalescent sera of COVID-19 patients and vaccine of BBV152, although the efficacy was low [64]. Detection of the SARS-CoV-2 viral genome, consisting of single-stranded RNA, is effectively done by reverse transcription quantitative polymerase chain reaction (RT-qPCR), which is the gold standard technique widely used in molecular diagnostics [65,66]. There are several practical considerations when performing diagnostic assays using RT-qPCR.…”

Section: Notable Variants Of Sars-cov-2mentioning

confidence: 99%

Nucleic Acid Testing of SARS-CoV-2

Yoo

Kim

2021

IJMS

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The coronavirus disease 2019 (COVID-19) has caused a large global outbreak. It is accordingly important to develop accurate and rapid diagnostic methods. The polymerase chain reaction (PCR)-based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA. Along with the RT-PCR method, digital PCR has emerged as a powerful tool to quantify nucleic acid of the virus with high accuracy and sensitivity. Non-PCR based techniques such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA) are considered to be rapid and simple nucleic acid detection methods and were reviewed in this paper. Non-conventional molecular diagnostic methods including next-generation sequencing (NGS), CRISPR-based assays and nanotechnology are improving the accuracy and sensitivity of COVID-19 diagnosis. In this review, we also focus on standardization of SARS-CoV-2 nucleic acid testing and the activity of the National Metrology Institutes (NMIs) and highlight resources such as reference materials (RM) that provide the values of specified properties. Finally, we summarize the useful resources for convenient COVID-19 molecular diagnostics.

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rRT-PCR for SARS-CoV-2: Analytical considerations

Cited by 71 publications

References 53 publications

Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Impact of the Nucleic Acid Extraction Method and the RT-qPCR Assay on SARS-CoV-2 Detection in Low-Viral Samples

Nucleic Acid Testing of SARS-CoV-2

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