A Nanoconfined, Hydrogel-Partitioned LAMP Platform for Rapid, Extraction-Free Quantification of Mycoplasma pneumoniae

IF 4.5 2区医学 Q2 CHEMISTRY, MEDICINAL

ACS Infectious Diseases Pub Date : 2026-07-10 Epub Date: 2026-06-12 DOI:10.1021/acsinfecdis.6c00203

Xiaoyang Jin, Yuxuan Li, Yajun Guo, Xingyu Lin*, Wei Li* and Danli Xie*,

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Abstract

Accurate quantification of Mycoplasma pneumoniae (M. pneumoniae) is important because pathogen load is associated with disease severity and treatment outcomes. However, quantitative polymerase chain reaction (qPCR) requires DNA extraction and standard-curve calibration, whereas digital polymerase chain reaction (dPCR) remains costly and less accessible. Here, we developed gLAMP-X, an extraction-free hydrogel-limited amplification (gLAMP) platform for rapid quantification of M. pneumoniae from clinical throat swabs. The “X” component denotes extraction-free lysis that disrupts the lipid bilayer of M. pneumoniae while preserving nucleic acid integrity. In gLAMP-X, a polyethylene glycol (PEG)-based nanoporous hydrogel partitions loop-mediated isothermal amplification (LAMP) into isolated nanoscopic microreactors, converting homogeneous amplification into a digital-like system in which each event forms a discrete fluorescent cluster for end point counting. Under optimized conditions, gLAMP-X achieved a limit of detection of 1.7 copies/μL within 20 min. In 90 pediatric throat swabs, gLAMP-X showed 100% sensitivity, 98.1% specificity, and strong quantitative agreement with qPCR.

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用于肺炎支原体快速、无提取定量的纳米限水凝胶分离LAMP平台。

准确定量肺炎支原体（肺炎支原体）是很重要的，因为病原体负荷与疾病严重程度和治疗结果相关。然而，定量聚合酶链反应（qPCR）需要DNA提取和标准曲线校准，而数字聚合酶链反应（dPCR）仍然昂贵且不易获得。在这里，我们开发了gLAMP- x，一种无提取水凝胶限制扩增（gLAMP）平台，用于快速定量临床咽拭子中的肺炎支原体。“X”成分表示无提取物裂解，破坏肺炎支原体的脂质双分子层，同时保持核酸的完整性。在gLAMP-X中，基于聚乙二醇（PEG）的纳米多孔水凝胶将环介导的等温扩增（LAMP）分割成孤立的纳米级微反应器，将均匀扩增转化为类似数字的系统，其中每个事件形成一个离散的荧光簇，用于端点计数。在优化条件下，gLAMP-X在20 min内的检出限为1.7 copies/μL。在90份儿童咽拭子中，gLAMP-X的灵敏度为100%，特异性为98.1%，与qPCR的定量结果高度一致。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Infectious Diseases CHEMISTRY, MEDICINALINFECTIOUS DISEASES&nb-INFECTIOUS DISEASES

CiteScore

9.70

自引率

3.80%

发文量

213

期刊介绍： ACS Infectious Diseases will be the first journal to highlight chemistry and its role in this multidisciplinary and collaborative research area. The journal will cover a diverse array of topics including, but not limited to: * Discovery and development of new antimicrobial agents — identified through target- or phenotypic-based approaches as well as compounds that induce synergy with antimicrobials. * Characterization and validation of drug target or pathways — use of single target and genome-wide knockdown and knockouts, biochemical studies, structural biology, new technologies to facilitate characterization and prioritization of potential drug targets. * Mechanism of drug resistance — fundamental research that advances our understanding of resistance; strategies to prevent resistance. * Mechanisms of action — use of genetic, metabolomic, and activity- and affinity-based protein profiling to elucidate the mechanism of action of clinical and experimental antimicrobial agents. * Host-pathogen interactions — tools for studying host-pathogen interactions, cellular biochemistry of hosts and pathogens, and molecular interactions of pathogens with host microbiota. * Small molecule vaccine adjuvants for infectious disease. * Viral and bacterial biochemistry and molecular biology.