ArticleClinica chimica acta; international journal of clinical chemistry2020
Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR.
Article in Clinica chimica acta; international journal of clinical chemistry, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 22 citations in OpenAlex.
- Article
- Integrated point-of-care RT-PCR methods during and after COVID-19 pandemic.Virusdisease · 2023Review
- Spike in pollution to ignite the bursting of COVID-19 second wave is more dangerous than spike of SAR-CoV-2 under environmental ignorance in long term: a review.Environmental science and pollution research international · 2022Review
- Rapid identification of SARS-CoV-2 in the point-of-care using digital PCR-based Dr. PCR™ Di20K COVID-19 Detection Kit without viral RNA extraction.Genes & genomics · 2022Article
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundsThe coronavirus disease 2019 (COVID-19) pandemic is still ongoing. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) is regarded as a gold-standard method for the diagnosis of COVID-19. However, unexpected contamination of synthesized positive control samples included in COVID-19 test kits have increased the inconclusiveness of disease interpretation. Therefore, it is important to establish new methods for the preparation of reliable positive controls that are not affected by contamination for the accurate for diagnosis of COVID-19, but it still remains a challenge.
methodsA new approach for producing synthetic positive controls using synthetic positive template (SPT) oligonucleotides was designed. SPT oligonucleotides contain probe binding and virus-irrelevant regions were used as templates for real-time PCR to evaluate the expression level of SARS-CoV-2 genes (RdRP, E, and N). The limit of detection (LOD) for individual SARS-CoV-2 genes by Ct values with different concentrations of SPT templates and genomic RNAs from SARS-CoV-2 infected samples was determined.
resultsLODs with SPT templates were >10
conclusionThis method significantly reduces the risk of false-positive reactions resulting from contamination in the synthesis procedures of positive control materials. Therefore, this approach could be integrated into the currrently available COVID-19 test kits and will provide a general method for preparing positive controls in the diagnosis of emerging RNA virus infections.
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