Multiplex Real-Time Reverse-Transcription Polymerase Chain Reaction Assays for Diagnostic Testing of Severe Acute Respiratory Syndrome Coronavirus 2 and Seasonal Influenza Viruses: A Challenge of the Phase 3 Pandemic Setting

Mancini, Fabiola; Barbanti, Fabrizio; Scaturro, Maria; Fontana, Stefano; Martino, Angela Di; Marsili, Giulia; Puzelli, Simona; Calzoletti, Laura; Facchini, Marzia; Mario, Giuseppina Di; Fabiani, Concetta; Bella, Antonino; Riccardo, Flavia; Pezzotti, Patrizio; Stefanelli, Paola; Rezza, Giovanni; Ciervo, Alessandra; Villa, Laura; Fortini, Daniela; Iacobino, Angelo; Fiore, Stefano; Benedetti, Eleonora; Marchi, Antonella; Venturi, Giulietta; Fortuna, Claudia; Amendola, Antonello; Toma, Luciano; Luca, Marco Di; Severini, Francesco

doi:10.1093/infdis/jiaa658

Cited by 25 publications

(27 citation statements)

References 20 publications

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“…Four set of primer/probes for E and RdRP genes of SARS-CoV-2, M gene of influenza A, and NS2 gene of influenza B were adapted and modified with 2′O-methylated RNA-bases at their penultimate (3′-end) positions to reduce formation of primer dimers [27]. In another study by Mancini et al, primers for influenza virus A and B, N2 and E genes of SARS-CoV-2, and the human RP gene as internal control, were used to simultaneously detect the viruses in the multiplex RT-PCR assay in 1000 clinical samples of which two coinfections of SARS-CoV-2 and influenzas were reported [28]. Moreover, the USA Centers for Disease Control and Prevention (CDC) developed the Influenza SARS-CoV-2 (Flu SC2) multiplex RT-PCR assay that was received Emergency Use Authorization (EUA) approval by FDA.…”

Section: Multiplex Real-time Pcr Technologymentioning

confidence: 99%

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

et al. 2021

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The ongoing pandemic of coronavirus disease 2019 (COVID-19) has made a serious public health and economic crisis worldwide which united global efforts to develop rapid, precise, and cost-efficient diagnostics, vaccines, and therapeutics. Numerous multi-disciplinary studies and techniques have been designed to investigate and develop various approaches to help frontline health workers, policymakers, and populations to overcome the disease. While these techniques have been reviewed within individual disciplines, it is now timely to provide a cross-disciplinary overview of novel diagnostic and therapeutic approaches summarizing complementary efforts across multiple fields of research and technology. Accordingly, we reviewed and summarized various advanced novel approaches used for diagnosis and treatment of COVID-19 to help researchers across diverse disciplines on their prioritization of resources for research and development and to give them better a picture of the latest techniques. These include artificial intelligence, nano-based, CRISPR-based, and mass spectrometry technologies as well as neutralizing factors and traditional medicines. We also reviewed new approaches for vaccine development and developed a dashboard to provide frequent updates on the current and future approved vaccines.

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Section: Multiplex Real-time Pcr Technologymentioning

confidence: 99%

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

et al. 2021

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“…We extracted the total RNA from the samples according to the manufacturer's instructions using the ExiStation™ and ExiPrep™ 48 Viral DNA/RNA Kit. We referred to the WHO protocol for the PCR conditions and primer sequences used for ampli cation [9]…”

Section: Sequencingmentioning

confidence: 99%

“…Moreover, simultaneous infection with SARS-CoV-2 and other respiratory pathogens may in uence the morbidity and prognosis in patients [8]. It is important to accurately detect SARS-CoV-2 and in uenza viruses, including possible coinfections in the community [9]. Reverse-transcription PCR (RT-PCR) is highly sensitive and speci c and has become the standard test for detecting respiratory viruses, including SARS-CoV-2 [8].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the AccuPower^® RV1 Real-Time RT-PCR Kit and the AccuPower^® RV1 Multiplex Kit for SARS-CoV-2 and Influenza Virus Detection

et al. 2021

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Background: The AccuPower ® RV1 Real-Time RT-PCR Kit (Bioneer, Korea) and AccuPower ® RV1 Multiplex Kit (Bioneer) are one-step real-time reverse transcription PCR assays for detecting severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza A and B. Methods: We clinically evaluated the AccuPower ® RV1 Real-Time RT-PCR Kit and AccuPower ® RV1 Multiplex Kit by comparing their results for 1,098 clinical samples. The presence of SARS-CoV-2 was confirmed using the Allplex™ 2019-nCoV Assay (Seegene, Korea) and Standard M nCoV Real-Time Detection Kit (SD Biosensor, Korea). Influenza viruses were detected using the Allplex™ Respiratory Panel 1 (Seegene). Results: The comparative positive and negative agreement values of the AccuPower ® RV1 Real-Time RT-PCR Kit for SARS-CoV-2 and influenza A and B were 100%. The positive agreement of the AccuPower ® RV1 Multiplex Kit was 100% for SARS-CoV-2 and 98.77% for influenza A and B. The kappa values for SARS-CoV-2 and influenza A and B were > 0.99. SARS-CoV-2 was evaluated using both sputum and nasopharyngeal or oropharyngeal swabs. There was no difference in the detection rates for each type. Conclusions: The findings confirm the clinically comparable performances of the AccuPower ® RV1 Real-Time RT-PCR Kit and the AccuPower ® RV1 Multiplex Kit.

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“…Several multiplex approaches have been developed for providing a quick molecular diagnosis of COVID-19 ( Attwood et al, 2020 ; Visseaux et al, 2020 ; Waggoner et al, 2020 ). Some multiplex protocols were developed for a dual detection of SARS-CoV-2 and influenza virus ( Chung et al, 2021 ; Mancini et al, 2020 ; Nörz et al, 2021 ), for the detection of multiple respiratory pathogens ( Hirotsu et al, 2020 ; Panning et al, 2020 ; Williams et al, 2020 ), and for the detection of variants ( Islam et al, 2021 ). Other multiplex approaches were aimed at library preparation for genome sequencing ( Paden et al, 2020 ; Tyson et al, 2020 ; Xiao et al, 2020 ) and respiratory viral infection trend analysis ( Marriott et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV-2 detection in wastewater using multiplex quantitative PCR

Navarro

Gómez

Sanseverino

et al. 2021

Science of The Total Environment

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A multiplex reverse transcription quantitative PCR (RT-qPCR)-based method was designed for the simultaneous detection of different SARS-CoV-2 genes. In this study, we used three target genes encoding for the nucleocapsid 1 and 3 (N1, N3), and the spike (S) proteins, all commonly used in the detection of SARS-CoV-2 in human and environmental samples. The performance of the multiplex assay, compared to the single assay was assessed for the standard calibration curve, required for absolute quantification, and then, for the real environmental samples to detect SARS-CoV-2. For this latter, four environmental samples were collected at a local wastewater treatment plant (WWTP). The results showed that the cycle threshold (Ct) values of the multiplex were comparable to the values obtained by the singleplex PCR. The amplification of the three target genes indicated the presence of SARS-CoV-2 in the four water samples with an increasing trend in February and these results were confirmed in the multiplex approach, showing the robustness of this method and its applicability for the relative abundance analysis among the samples. Overall, both the laboratory and field work results demonstrated that the multiplex PCR assay developed in this study could provide a method for SARS-CoV-2 detection as robust as the single qPCR, but faster and cost-effective, reducing by three times the number of reactions, and consequently the handling time and reagents.

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Multiplex Real-Time Reverse-Transcription Polymerase Chain Reaction Assays for Diagnostic Testing of Severe Acute Respiratory Syndrome Coronavirus 2 and Seasonal Influenza Viruses: A Challenge of the Phase 3 Pandemic Setting

Cited by 25 publications

References 20 publications

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

Evaluation of the AccuPower^® RV1 Real-Time RT-PCR Kit and the AccuPower^® RV1 Multiplex Kit for SARS-CoV-2 and Influenza Virus Detection

SARS-CoV-2 detection in wastewater using multiplex quantitative PCR

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Multiplex Real-Time Reverse-Transcription Polymerase Chain Reaction Assays for Diagnostic Testing of Severe Acute Respiratory Syndrome Coronavirus 2 and Seasonal Influenza Viruses: A Challenge of the Phase 3 Pandemic Setting

Cited by 25 publications

References 20 publications

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

Evaluation of the AccuPower® RV1 Real-Time RT-PCR Kit and the AccuPower® RV1 Multiplex Kit for SARS-CoV-2 and Influenza Virus Detection

SARS-CoV-2 detection in wastewater using multiplex quantitative PCR

Contact Info

Product

Resources

About

Evaluation of the AccuPower^® RV1 Real-Time RT-PCR Kit and the AccuPower^® RV1 Multiplex Kit for SARS-CoV-2 and Influenza Virus Detection