Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection

Alcoba-Flórez, Julia; Gil-Campesino, Helena; Artola, Diego García-Martínez de; González-Montelongo, Rafaela; Valenzuela-Fernández, Agustı́n; Ciuffreda, Laura; Flores, Carlos

doi:10.1016/j.ijid.2020.07.058

Cited by 58 publications

(34 citation statements)

References 11 publications

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“…The genome of SARS-CoV-2 consists of another type of nucleic acid, ribonucleic acid (RNA), which is similar in chemical composition to DNA, but has a number of fundamental differences. DNA stores hereditary information, and RNA transfer it; furthermore, DNA is a double-stranded molecule and is located in the nucleus, while RNA is single-stranded, located in cytoplasm, and has a lower molecular weight [ 32 , 36 , 37 , 38 ].…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

“…In addition to the qualitative determination of RNA in viral infections, a technology combined with qPCR to quantify the expressed gene is also often used. This combined option, referred to as real-time quantitative RT-PCR (qRT-PCR or RT-qPCR), is the most informative and accurate modification of the “classic” PCR for detection and quantification of RNA level [ 33 , 35 , 38 ]. This method is quantitative (or semi-quantitative)—during the PCR process, the sample is constantly monitored by a sensor that records fluorescent signals at each cycle of the reaction.…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

“…However, as often happens in clinical laboratory diagnostics, the importance of the preanalytical stage is overlooked, including collection and transportation of biomaterial, which, as a rule, is performed by untrained personnel or even by patients themselves (self-sampling, self-swab tests). This reduces confidence in results, especially if one uses screening assay [ 26 , 27 , 31 , 38 , 40 , 42 ].…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

“…Sampling with cotton swabs can wash out epithelial cells and reduce nucleic acid extraction. These samples are easier to collect, especially in resource-limited conditions [ 20 , 21 , 29 , 38 , 45 ]. It is extremely important to properly collect material in order to minimize the false-negative rate among COVID-19 positive patients.…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

“…Samples should be stored at 2–8 °C for up to 72 h [ 11 , 35 , 36 ]. If longer storage is required, samples are frozen to −70 °C and below [ 35 , 37 , 38 , 39 ].…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

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Laboratory-Based Resources for COVID-19 Diagnostics: Traditional Tools and Novel Technologies. A Perspective of Personalized Medicine

Andryukov

Беседнова

Кузнецова

et al. 2021

JPM

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The coronavirus infection 2019 (COVID-19) pandemic, caused by the highly contagious SARS-CoV-2 virus, has provoked a global healthcare and economic crisis. The control over the spread of the disease requires an efficient and scalable laboratory-based strategy for testing the population based on multiple platforms to provide rapid and accurate diagnosis. With the onset of the pandemic, the reverse transcription polymerase chain reaction (RT-PCR) method has become a standard diagnostic tool, which has received wide clinical use. In large-scale and repeated examinations, these tests can identify infected patients with COVID-19, with their accuracy, however, dependent on many factors, while the entire process takes up to 6–8 h. Here we also describe a number of serological systems for detecting antibodies against SARS-CoV-2. These are used to assess the level of population immunity in various categories of people, as well as for retrospective diagnosis of asymptomatic and mild COVID-19 in patients. However, the widespread use of traditional diagnostic tools in the context of the rapid spread of COVID-19 is hampered by a number of limitations. Therefore, the sharp increase in the number of patients with COVID-19 necessitates creation of new rapid, inexpensive, sensitive, and specific tests. In this regard, we focus on new laboratory technologies such as loop mediated isothermal amplification (LAMP) and lateral flow immunoassay (LFIA), which have proven to work well in the COVID-19 diagnostics and can become a worthy alternative to traditional laboratory-based diagnostics resources. To cope with the COVID-19 pandemic, the healthcare system requires a combination of various types of laboratory diagnostic testing techniques, whodse sensitivity and specificity increases with the progress in the SARS-CoV-2 research. The testing strategy should be designed in such a way to provide, depending on the timing of examination and the severity of the infection in patients, large-scale and repeated examinations based on the principle: screening–monitoring–control. The search and development of new methods for rapid diagnostics of COVID-19 in laboratory, based on new analytical platforms, is still a highly important and urgent healthcare issue. In the final part of the review, special emphasis is made on the relevance of the concept of personalized medicine to combat the COVID-19 pandemic in the light of the recent studies carried out to identify the causes of variation in individual susceptibility to SARS-CoV-2 and increase the efficiency and cost-effectiveness of treatment.

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Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

“…Samples should be stored at 2–8 °C for up to 72 h [ 11 , 35 , 36 ]. If longer storage is required, samples are frozen to −70 °C and below [ 35 , 37 , 38 , 39 ].…”

Section: Molecular Technologies For Identification Of Nucleic Acidmentioning

confidence: 99%

See 3 more Smart Citations

Laboratory-Based Resources for COVID-19 Diagnostics: Traditional Tools and Novel Technologies. A Perspective of Personalized Medicine

Andryukov

Беседнова

Кузнецова

et al. 2021

JPM

View full text Add to dashboard Cite

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A Versatile Biomimic Nanotemplating Fluidic Assay for Multiplex Quantitative Monitoring of Viral Respiratory Infections and Immune Responses in Saliva and Blood

et al. 2022

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The last pandemic exposed critical gaps in monitoring and mitigating the spread of viral respiratory infections at the point‐of‐need. A cost‐effective multiplexed fluidic device (NFluidEX), as a home‐test kit analogous to a glucometer, that uses saliva and blood for parallel quantitative detection of viral infection and body's immune response in an automated manner within 11 min is proposed. The technology integrates a versatile biomimetic receptor based on molecularly imprinted polymers in a core–shell structure with nano gold electrodes, a multiplexed fluidic‐impedimetric readout, built‐in saliva collection/preparation, and smartphone‐enabled data acquisition and interpretation. NFluidEX is validated with Influenza A H1N1 and SARS‐CoV‐2 (original strain and variants of concern), and achieves low detection limit in saliva and blood for the viral proteins and the anti‐receptor binding domain (RBD) Immunoglobulin G (IgG) and Immunoglobulin M (IgM), respectively. It is demonstrated that nanoprotrusions of gold electrodes are essential for the fine templating of antibodies and spike proteins during molecular imprinting, and differentiation of IgG and IgM in whole blood. In the clinical setting, NFluidEX achieves 100% sensitivity and 100% specificity by testing 44 COVID‐positive and 25 COVID‐negative saliva and blood samples on par with the real‐time quantitative polymerase chain reaction (p < 0.001, 95% confidence) and the enzyme‐linked immunosorbent assay.

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Pediatric Ischemic Stroke: An Infrequent Complication of SARS‐CoV‐2

Beslow

Linds

Fox

et al. 2021

Annals of Neurology

106

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Objective Severe complications of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) include arterial ischemic stroke (AIS) in adults and multisystem inflammatory syndrome in children. Whether stroke is a frequent complication of pediatric SARS‐CoV‐2 is unknown. This study aimed to determine the proportion of pediatric SARS‐CoV‐2 cases with ischemic stroke and the proportion of incident pediatric strokes with SARS‐CoV‐2 in the first 3 months of the pandemic in an international cohort. Methods We surveyed 61 international sites with pediatric stroke expertise. Survey questions included: numbers of hospitalized pediatric (≤ 18 years) patients with SARS‐CoV‐2; numbers of incident neonatal and childhood ischemic strokes; frequency of SARS‐CoV‐2 testing for pediatric patients with stroke; and numbers of stroke cases positive for SARS‐CoV‐2 from March 1 to May 31, 2020. Results Of 42 centers with SARS‐CoV‐2 hospitalization numbers, 8 of 971 (0.82%) pediatric patients with SARS‐CoV‐2 had ischemic strokes. Proportions of stroke cases positive for SARS‐CoV‐2 from March to May 2020 were: 1 of 108 with neonatal AIS (0.9%), 0 of 33 with neonatal cerebral sinovenous thrombosis (CSVT; 0%), 6 of 166 with childhood AIS (3.6%), and 1 of 54 with childhood CSVT (1.9%). However, only 30.5% of neonates and 60% of children with strokes were tested for SARS‐CoV‐2. Therefore, these proportions represent 2.9, 0, 6.1, and 3.0% of stroke cases tested for SARS‐CoV‐2. Seven of 8 patients with SARS‐CoV‐2 had additional established stroke risk factors. Interpretation As in adults, pediatric stroke is an infrequent complication of SARS‐CoV‐2, and SARS‐CoV‐2 was detected in only 4.6% of pediatric patients with ischemic stroke tested for the virus. However, < 50% of strokes were tested. To understand the role of SARS‐CoV‐2 in pediatric stroke better, SARS‐CoV‐2 testing should be considered in pediatric patients with stroke as the pandemic continues. ANN NEUROL 2021;89:657–665

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Sensitivity of different RT-qPCR solutions for SARS-CoV-2 detection

Abstract: Highlights This study compared RT-qPCR sensitivity for SARS-CoV-2 detection. False negatives were in the range of 2–39.8%. The most sensitive solution (97.9% [92.8–99.7]) targeted the SARS-CoV-2 E-gene.

Cited by 58 publications

References 11 publications

Laboratory-Based Resources for COVID-19 Diagnostics: Traditional Tools and Novel Technologies. A Perspective of Personalized Medicine

Laboratory-Based Resources for COVID-19 Diagnostics: Traditional Tools and Novel Technologies. A Perspective of Personalized Medicine

A Versatile Biomimic Nanotemplating Fluidic Assay for Multiplex Quantitative Monitoring of Viral Respiratory Infections and Immune Responses in Saliva and Blood

Pediatric Ischemic Stroke: An Infrequent Complication of SARS‐CoV‐2

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