Polyethylene glycol modified LAMP assay enables sensitive and specific clinical detection of varicella-zoster virus
Accurate, timely diagnosis of varicella-zoster virus (VZV) is important for treatment and infection control. While loop-mediated isothermal amplification (LAMP) is operationally simple, nonspecific priming can degrade performance. We assessed a polyethylene glycol-modified LAMP (PEG-LAMP) that tunes the reaction microenvironment via macromolecular crowding. PEG (1 μL per reaction) was titrated across concentrations; 100 mM was selected as the optimized condition because negatives remained at baseline while target amplification kinetics were maintained. PEG-LAMP preserved a log-linear relation between threshold time and input and improved the detection limit from 10<sup>3</sup> to 10<sup>2</sup> copies/μL compared with conventional LAMP. Precision at a fixed input exhibited low variability, and specificity was supported by flat traces in non-target reactions. In a 30-sample panel (15 spiked positives, 15 negatives) tested in parallel by PCR, conventional LAMP, and PEG-LAMP, PEG-LAMP was fully concordant with PCR and yielded shorter time-to-threshold for positives, whereas conventional LAMP produced one false negative and four false positives. Taken together, the results demonstrate that microenvironmental tuning with PEG provides a low-complexity means to suppress nonspecific LAMP while preserving on-target amplification, yielding a lower detection limit and faster time-to-result with PCR-level qualitative agreement in clinical VZV diagnosis.
Authors
- Wei Guo (ORCID: https://orcid.org/0000-0003-4649-1047)
- Bin Xu (ORCID: https://orcid.org/0000-0001-5129-9303)
- Li Yang (ORCID: https://orcid.org/0000-0001-6723-7377)
- Lei Zhang (ORCID: https://orcid.org/0000-0002-0083-6583)
- Wenjie Fang (ORCID: https://orcid.org/0000-0003-2936-6391)
- Shengcai Lv
- Jun Tian (ORCID: https://orcid.org/0009-0009-2756-6366)
Institutions
- Shanxi Medical University (CN)
- Shaanxi Provincial People's Hospital (CN)
- First Affiliated Hospital of Jiangxi Medical College (CN)
- Jiangxi Provincial Cancer Hospital (CN)
- Shanghai Changzheng Hospital (CN)
- Xijing Hospital (CN)
- Shanxi Provincial People’s Hospital (CN)
- Air Force Medical University (CN)
Publication Details
- Journal
- Frontiers in Cellular and Infection Microbiology
- Published
- 2025-12-09
- DOI
- https://doi.org/10.3389/fcimb.2025.1718986
- Primary Topic
- Biosensors and Analytical Detection
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Shaanxi Administration of Traditional Chinese Medicine
- Ministry of Education, India