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

Kim, Man Jin; Park, Hyunwoong; Jeon, You La; Shin, Ho Seob; Cho, Sung Im; Kim, Boram; Lee, Jee-Soo; Park, Sung Sup; Seong, Moon Woo

doi:10.47429/lmo.2021.11.4.290

Cited by 2 publications

References 16 publications

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Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Abbasi,

Nikoo,

Fotouhi

et al. 2023

BMC Microbiol

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Background During the coronavirus disease 2019 (COVID-19) pandemic, the simultaneous detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Influenza A, and Influenza B viruses is essential for rapid differential diagnosis in patients with similar symptoms, especially during “flu season” in the post-pandemic era. So far, several multiplex methods have been approved for the simultaneous detection of SARS-CoV-2, Influenza A, and Influenza B. However, due to the rapid mutation rate of the SARS-CoV-2 genome and the emergence of new variants, existing methods must be improved and updated. Methods To identify a highly conserved region in the SARS-CoV-2 N-gene, a genomic survey was performed to increase the sensitivity and specificity of primer and probe sets targeting the SARS-CoV-2 genome. The 95% LLOD (95% lower limits of detection) were calculated by probit analysis. A total of 70 predetermined clinical samples using singleplex RT-qPCR assays, were included. The clinical performance of the multiplex RT-qPCR assay was determined and compared with a commercial multiplex kit. The Cohen’s kappa coefficient, P-value (McNemar’s test), Passing-Bablok regression, and Bland Altman agreement analysis were determined to monitor the agreement of the assays. Results The novel SARS-CoV-2 primer and probe set designed in this assay was able to detect all variants of concern (VOCs) and variants of interest (VOIs) with high analytical and clinical performance. The 95% LLOD for the multiplex RT-qPCR was 20 copies per reaction for the N gene of SARS-CoV-2, 2 copies per reaction for M1 gene of Influenza A and NS1 gene of Influenza B. The diagnostic sensitivity of the multiplex RT-qPCR was 94.4%, 93.7%, and 100% for the detection of SARS-CoV-2, Influenza A, and Influenza B genomes, respectively. Moreover, the specificity was identical (100%) in both assays. According to the agreement analysis results, there was no statistical difference between our multiplex assay and the commercial kit. Conclusions In this study, we developed a novel in-house made multiplex RT-qPCR assay, with high sensitivity, specificity, and reliability for the diagnosis of SARS-CoV-2 infection in clinical samples. This is valuable during Influenza seasons when influenza co-circulates with SARS-CoV-2, as it saves costs, time, and thus specific and timely treatment of patients.

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Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Abbasi,

Nikoo,

Fotouhi

et al. 2023

BMC Microbiol

View full text Add to dashboard Cite

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Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for Simultaneous detection of SARS-CoV-2 and influenza A/B viruses

Abbasi

Nikoo

Fotouhi

et al. 2023

Preprint

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Background: During the coronavirus disease 2019 (COVID-19) pandemic, the simultaneous detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A/B viruses are essential in the rapid differential diagnosis in patients with similar symptoms. So far, several multiplex methods were published for simultaneous detection of SARS-CoV-2 and Flu A/B. However, due to the rapid mutation rate of the SARS-CoV-2 genome and the emergence of new variants, making it necessary to improve and update existing methods. Methods: To identify a highly conserved region on SARS-CoV-2 N-gene, variant genome survey was performed to increase the sensitivity and specificity of primers and probe annealing to the SARS-CoV-2 genome. The analytical and diagnostic sensitivity and specificity of our multiplex RT-qPCR assay were determined; and the kappa agreement was calculated to highlight the agreement using the Chi-square test by SPSS 22.0. Results: The new set of SARS-CoV-2 primer-probes designed in the present assay was able to detect all variants of concern (VOCs) and variants of interest (VOIs) with high analytical specificity among other respiratory viruses. The analytical sensitivity by our multiplex assay was ≤ 30 copies of the plasmid per reaction. A total of 70 clinical samples were analyzed using our multiplex assay and compared with multiplex commercial kit. The sensitivity of our multiplex RT-qPCR assay and commercial kit were 95.4% and 93.1%, respectively. Also, the specificity was the same (100%) in both methods. Kappa agreement was measured at 0.97 and 0.90 for our multiplex assay and commercial kit respectively. Conclusions: A novel and robust TaqMan probe-based one-step multiplex RT-qPCR assay, with high analytical and clinical performance and agreement was developed and validated in this study. It will be valuable in influenza seasons when influenza expected co-circulate with SARS-CoV-2, due to saving costs, time, and consequently, specific and timely treatment of patients.

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

Cited by 2 publications

References 16 publications

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for Simultaneous detection of SARS-CoV-2 and influenza A/B viruses

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

Cited by 2 publications

References 16 publications

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses

Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for Simultaneous detection of SARS-CoV-2 and influenza A/B viruses

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