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.

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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

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article

Polyethylene glycol modified LAMP assay enables sensitive and specific clinical detection of varicella-zoster virus

Wei Guo, Bin Xu, Li Yang, Lei Zhang et al.
Frontiers in Cellular and Infection Microbiology
Biosensors and Analytical Detection
article

Polyethylene glycol modified LAMP assay enables sensitive and specific clinical detection of varicella-zoster virus

Wei Guo, Bin Xu, Li Yang, Lei Zhang, Wenjie Fang, Shengcai Lv, Jun Tian
article en

Abstract

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.

Frontiers in Cellular and Infection MicrobiologyVol. 15
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)
Shaanxi Administration of Traditional Chinese Medicine, Ministry of Education, India
Openalex Percentile: Top 62%
Biosensors and Analytical Detection
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