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2025 Vol.55, Issue 2 Preview Page

Original Article

Development of a Multiplex One-step RT-qPCR Diagnostic Test for SARS-CoV-2 Using a Portable Biomeme Thermocycler
Seonghyeon Lee1†
,
Shivani Rajoriya1†
,
Jieun Park1
,
Bohyeon Kim1
,
Augustine Natasha1
,
Sara Puspareni Prayitno1
,
Divya Misra1
,
Kyungmin Park23
,
Jin-Won Song23
,
Taek-Kyun Lee4
,
Kyu Sung Shin5
,
Hyungdon Lee6
,
Won-Keun Kim17*
1Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252, Republic of Korea
2Department of Microbiology, Korea University College of Medicine, Seoul 02841, Republic of Korea
3BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul 02841, Republic of Korea
4South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
5Department of Laboratory Medicine, Hallym University Chuncheon Sacred Heart Hospital, Hallym University College of Medicine, Chuncheon 24253, Republic of Korea
6Division of Infectious Diseases, Department of Internal Medicine, Hallym University Chuncheon Sacred Heart Hospital, Hallym University College of Medicine, Chuncheon 24253, Republic of Korea
7Institute of Medical Research, College of Medicine, Hallym University, Chuncheon 24252, Republic of Korea
*wkkim1061@hallym.ac.kr
†These authors equally contributed to this work.
30 June 2025. pp. 142-159
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Abstract

Coronavirus disease-19 (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), poses a critical public health threat due to its high transmissibility, infectivity, and prolonged incubation in humans. Timely identification of SARS-CoV-2 is crucial to control COVID-19 in the lack of effective antiviral medications in low resource regions. However, an accurate assay and portable diagnostic method for SARS-CoV-2 remains to be developed. To address this issue, we developed a multiplex one-step reverse transcription quantitative polymerase chain reaction (RT-qPCR) for diagnosis of SARS-CoV-2 using a portable Biomeme Franklin™ Real-Time PCR thermocycler. The performance of the Biomeme assay was evaluated and validated for the testing of SARS-CoV-2 based on multiplex detection of the nucleocapsid, envelope, and spike genes. The Biomeme assay demonstrated high sensitivity by detecting the RNA of wild type 19 A and multiple SARS-CoV-2 variants, including B.1.617.2, BA.1, BA.2, BA.2.75, BA.5, and BN.1 from 0.01 ng/µl of total RNA and showed no cross-reactivity with other human coronaviruses, such as HCoV-OC43, HCoV-NL63, and HCoV-229E. The diagnostic method exhibited an analytical sensitivity of 95% and a specificity of 100%. The Biomeme Franklin platform generates rapid results within 3-4 h and allows smartphone-based output quantification, making it highly suitable as a Point of Care (POC) diagnostic tool. Therefore, this study reports on a diagnostic testing method for the prompt and reliable molecular diagnosis of SARS-CoV-2.

Keywords
SARS-CoV-2
Biomeme
Diagnostic
One step RT-qPCR
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Information
  • Publisher :The Korean Society for Microbiology and The Korean Society of Virology
  • Publisher(Ko) :대한미생물학회‧대한바이러스학회
  • Journal Title :JOURNAL OF BACTERIOLOGY AND VIROLOGY
  • Volume : 55
  • No :2
  • Pages :142-159
  • Received Date : 2025-04-24
  • Revised Date : 2025-06-04
  • Accepted Date : 2025-06-09
  • DOI :https://doi.org/10.4167/jbv.2025.55.2.142
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