Sun, Y.; Tang, D.; Li, N.; Wang, Y.; Yang, M.; Shen, C. Development of a Rapid Epstein–Barr Virus Detection System Based on Recombinase Polymerase Amplification and a Lateral Flow Assay. Viruses2024, 16, 106.
Sun, Y.; Tang, D.; Li, N.; Wang, Y.; Yang, M.; Shen, C. Development of a Rapid Epstein–Barr Virus Detection System Based on Recombinase Polymerase Amplification and a Lateral Flow Assay. Viruses 2024, 16, 106.
Sun, Y.; Tang, D.; Li, N.; Wang, Y.; Yang, M.; Shen, C. Development of a Rapid Epstein–Barr Virus Detection System Based on Recombinase Polymerase Amplification and a Lateral Flow Assay. Viruses2024, 16, 106.
Sun, Y.; Tang, D.; Li, N.; Wang, Y.; Yang, M.; Shen, C. Development of a Rapid Epstein–Barr Virus Detection System Based on Recombinase Polymerase Amplification and a Lateral Flow Assay. Viruses 2024, 16, 106.
Abstract
The quality of cellular products used in biological research can directly impact the ability to ob-tain accurate results. Epstein-Barr virus (EBV) is a latent virus that spreads extensively world-wide, and cell lines used in experiments may carry EBV and pose an infection risk. The presence of EBV in a single cell line can contaminate other cell lines used in the same laboratory, affecting experimental results. We developed three EBV detection systems: (1) a polymerase chain reac-tion (PCR)-based detection system, (2) a recombinase polymerase amplification (RPA)-based de-tection system, and (3) a combined RPA-lateral flow assay (LFA) detection system. The mini-mum EBV detection limits were 1×103 copy numbers for the RPA-based and RPA-LFA systems and 1×104 copy numbers for the PCR-based system. Both the PCR and RPA detection systems were applied to 192 cell lines, and the results were consistent with those obtained by the EBV assay methods specified in the pharma-ceutical industry standards of the People's Republic of China. 10 EBV-positive cell lines were identified. The combined RPA-LFA system is simple to operate, allowing for rapid results visualization. This system can be implemented in laborato-ries and cell banks as part of a daily quality control strategy to ensure cell quality and experi-mental safety and may represent a potential new technique for the rapid detection of EBV in clinical samples.
Keywords
RPA; LFA; EBV; Cell quality control
Subject
Biology and Life Sciences, Virology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
