key: cord-0800134-0etljojf
authors: Kim, Tae Yeul; Kim, Ji-Youn; Shim, Hyang Jin; Yun, Sun Ae; Jang, Ja-Hyun; Huh, Hee Jae; Kim, Jong-Won; Lee, Nam Yong
title: Comparison of the PowerChek SARS-CoV-2, Influenza A&B, RSV Multiplex Real-time PCR Kit and BioFire Respiratory Panel 2.1 for simultaneous detection of SARS-CoV-2, influenza A and B, and respiratory syncytial virus
date: 2021-09-28
journal: J Virol Methods
DOI: 10.1016/j.jviromet.2021.114304
sha: 7f3000300c983b70c97d316caf2888a670b560b3
doc_id: 800134
cord_uid: 0etljojf

The potential co-circulation of SARS-CoV-2, influenza, and respiratory syncytial virus (RSV) could pose an unprecedented challenge to healthcare systems worldwide. Here, we compared the performance of the PowerChek SARS-CoV-2, Influenza A&B, RSV Multiplex Real-time PCR Kit (PowerChek) for simultaneous detection of SARS-CoV-2, influenza A and B, and respiratory syncytial virus with that of BioFire Respiratory Panel 2.1 (RP2.1) using 175 nasopharyngeal swab (NPS) specimens. Positive percent agreement and negative percent agreement of the PowerChek assay compared to RP2.1 were as follows: 100% (40/40) and 100% (135/135) for SARS-CoV-2; 100% (39/39) and 100% (136/136) for influenza A; 100% (35/35) and 100% (140/140) for influenza B; and 93.1% (27/29) and 100% (146/146) for RSV, respectively. The limit of detection (LOD) was accessed using RNA standards for each virus, and the LOD values of the PowerChek assay for SARS-CoV-2, influenza A and B, and RSV were 0.36, 1.24, 0.09, and 0.63 copies/μL, respectively. Our results demonstrate that the PowerChek assay is sensitive and accurate for detection of SARS-CoV-2, influenza A and B, and RSV, suggesting that this assay can be a valuable diagnostic tool when SARS-CoV-2, influenza, and RSV are co-circulating.

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PowerChek; SARS-CoV-2; COVID-19; influenza; RSV; real-time PCR Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread rapidly, resulting in characterization of this outbreak as a pandemic. During this era of the SARS-CoV-2 pandemic, co-circulation of influenza and respiratory syncytial virus (RSV) could pose a challenge to healthcare providers as these viral infections have overlapping clinical presentations Solomon et al., 2020; Zayet et al., 2020) . Further complicating the situation is that co-infection of SARS-CoV-2 and other respiratory viruses, including influenza and RSV, is possible (Burrel et al., 2021; Cuadrado-Payan et al., 2020; Ding et al., 2020; Kim et al., 2020; Ma et al., 2020; Wu et al., 2020; Zhang et al., 2020) ; therefore, Table 1 ). All specimens were stored at −70°C until tested using the PowerChek and RP2.1 assays.

RNA was extracted from NPS specimens using the QIAamp DSP Viral RNA Mini Kit AmpliRun Influenza A H1, Influenza B, and RSV subtype A RNA Control [Vircell, Granada, Spain]) were used for analytical sensitivity evaluation. These RNA standards were serially diluted, and multiple replicates of each dilution were tested using the PowerChek assay. The limit of detection (LOD) was calculated using Probit regression analysis. Analytical specificity was evaluated using 20 respiratory virus strains (Table 1) .

For SARS-CoV-2, the PPA and NPA between the PowerChek assay and RP2.1 were 100% (40/40) and 100% (135/135), respectively ( (SARS-CoV-2 and influenza A and B), which suggests almost perfect agreement. Two specimens produced discordant results between the PowerChek assay and RP2.1 for RSV J o u r n a l P r e -p r o o f (Table 3) . They were PowerChek-negative and RP2.1-positive for RSV. After discrepancy resolution, one specimen was confirmed as positive for RSV, and this sample's high Ct value (39.5) indicates a low RSV viral load in the specimen. The LOD values of the PowerChek assay for SARS-CoV-2, influenza A and B, and RSV were 0.36, 1.24, 0.09, and 0.63 copies/μL, respectively (Table 4) , which were comparable to or higher than the claimed LOD values of the RP2.1 assay in the package insert (SARS-CoV-2: 0.5 copies/μL for heatinactivated virus and 0.16 copies/μL for infectious virus; influenza A H1: 0.14 copies/μL; influenza B: 0.034 copies/μL; RSV: 0.009 copies/μL). In the analytical specificity study, the

PowerChek assay detected only its intended targets (SARS-CoV-2, influenza A and B, and RSV) and showed no cross-reactivity with other respiratory viruses (Table 1) .

Currently, several multiplex rRT-PCR assays for simultaneous detection of respiratory viruses including SARS-CoV-2 are commercially available (Chung et al., 2021; Creager et al., 2020; Eckbo et al., 2021; Jarrett et al., 2021; Leung et al., 2021; Mostafa et al., 2020; Visseaux et al., 2020) . Most of these assays have been developed by adding SARS-CoV-2 testing to existing multiplex assays for detection of other respiratory viruses including influenza and RSV. The RP2.1, the Xpert Xpress SARS-CoV-2/Flu/RSV (Cepheid, Sunnyvale, CA, USA), the QIAstat-Dx respiratory SARS-CoV-2 panel (Qiagen, Hilden, Germany), and the ePlex Respiratory Pathogen Panel 2 (GenMark Diagnostics, Carlsbad, CA, USA) are such assays, and their performance has been assessed in previous studies (Creager et al., 2020; Eckbo et al., 2021; Jarrett et al., 2021; Leung et al., 2021; Mostafa et al., 2020; Visseaux et al., 2020) . Although these random-access assays make test results available to clinicians in a timely manner, they have a relatively low, albeit scalable, throughput and might not be suitable for high-volume laboratories. On the other hand, the

PowerChek assay is a high-throughput batch testing, suitable for laboratories performing a large number of assays.

Limitations of this single-center study are its retrospective design and small sample size.

A prospective study was not feasible as influenza-and RSV-positive samples have rarely been found in our hospital during the SARS-CoV-2 pandemic. Therefore, stored clinical specimens were selectively included to evaluate the performance of the PowerChek assay.

According to our study, the performance of the PowerChek assay was comparable to that of the RP2.1 assay in detecting SARS-CoV-2, influenza A and B, and RSV. Our results indicate that the PowerChek assay is a useful diagnostic tool for simultaneous detection of SARS-CoV-2, influenza, and RSV. 

The study protocol was reviewed and approved by the Institutional Review Board of Samsung Medical Center (IRB no. SMC 2020-12-061). 

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

None of the authors have competing interests to declare.