Clade-specific multiplex digital PCR assay for monkeypox virus detection in spike-in wastewater samples.

Access microbiology Pub Date : 2026-08-20 eCollection Date: 2026-01-01 DOI:10.1099/acmi.0.001041.v3
Melissa Pitton, Charles Gan, Patrick Schmidhalter, Christoph Ort, Timothy R Julian
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引用次数: 0

Abstract

Monkeypox virus (MPXV) is a zoonotic pathogen that has recently caused outbreaks in non-endemic areas. Wastewater-based surveillance is independent of individual testing and offers substantial potential for monitoring population-level disease dynamics of MPXV at the clade level, as clades differ in transmissibility and mortality rate. Here, we report the validation of a four-plex digital PCR assay to detect and quantify the major clades and subclades of MPXV. The assay demonstrated strong linearity, analytical sensitivity (limits of detection ranged from 6.7 to 9.6 copies per reaction across targets) and specificity in distinguishing and quantifying MPXV by clade, which is advantageous for both clinical and wastewater applications.

钉钉厂废水样品中猴痘病毒检测的分支特异性多重数字PCR方法。
猴痘病毒(MPXV)是一种人畜共患病原体,最近在非流行地区引起暴发。基于废水的监测独立于个体检测,具有在支系水平上监测MPXV种群水平疾病动态的巨大潜力,因为支系在传播性和死亡率方面存在差异。在这里,我们报告了一种四重数字PCR检测和量化MPXV主要分支和亚分支的验证。该方法具有很强的线性、分析灵敏度(检测限为6.7 ~ 9.6个拷贝)和特异性,可根据进化支区分和定量MPXV,这对临床和废水应用都是有利的。
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CiteScore
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