Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand

IF 1.9 Q2 MULTIDISCIPLINARY SCIENCES

MethodsX Pub Date : 2025-08-15 DOI:10.1016/j.mex.2025.103568

Wansadaj Jaroenram , Yada Ongchanchai , Ravee Nitiyanontakij , Pawita Suwanwatthana , Ratchana Ponang , Wansika Kiatpathomchai

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Abstract

Despite numerous recent advances in infectious disease diagnostic technologies including quantitative (q) polymerase chain reaction (PCR) and antigen test kit (ATK), there remains a need to develop more efficient point-of-care assays for large-scale operations. In an effort towards such a screening test for low-resource settings, we have established a colorimetric loop-mediated isothermal amplification assay and detected the amplified DNA via naked-eye observation to maximize assay simplicity using monkeypox (Mpox) as a model. The assay is cheap, highly sensitive and specific to Mpox with the potential for translation into broader applications.

•
Our protocol offers a simpler, faster, and cheaper test for Mpox than the current gold-standard qPCR, with 100% accuracy.
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assay enables decentralized screening and provides visually interpretable results immediately after incubation
•
The versatility of this protocol allows for easy adaptation to detecting new or re-emerging diseases, utilizing a visual readout for maximum simplicity.

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一种用于m痘诊断的目视LAMP测定法的验证和应用——来自泰国临床样本的见解

尽管最近在传染病诊断技术方面取得了许多进展，包括定量(q)聚合酶链反应（PCR）和抗原检测试剂盒（ATK），但仍需要为大规模手术开发更有效的即时检测方法。为了在低资源环境中进行这样的筛选试验，我们建立了比色环介导的等温扩增试验，并通过裸眼观察检测扩增的DNA，以猴痘（Mpox）为模型，最大限度地提高了试验的简洁性。该检测方法价格低廉，对m痘具有高度敏感性和特异性，具有转化为更广泛应用的潜力。•我们的方案提供了一个更简单，更快，更便宜的Mpox测试比目前的金标准qPCR，具有100%的准确性。•该方案的多功能性允许轻松适应检测新的或重新出现的疾病，利用视觉读数来最大限度地简化。

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