A Sensitive Real-Time PCR-Based Assay for the Identification of Colletotrichum in Phytosanitary and Clinical Applications

TL;DR

This paper introduces a fast and sensitive PCR test for detecting Colletotrichum fungi in both plant and human health contexts.

Contribution

A novel real-time PCR assay targeting the 28S rDNA region for universal and sensitive detection of Colletotrichum species.

Findings

The assay achieved a detection limit of 680 fg of genomic DNA.

It showed no cross-reactivity with related fungal species.

The method successfully identified Colletotrichum in complex environmental samples.

Abstract

Colletotrichum species are major plant pathogens and emerging opportunistic human pathogens. Due to their vast genetic diversity, existing diagnostic tools often suffer from narrow specificity or labor-intensive workflows. In this study, we developed a rapid, universal, and highly sensitive genus-specific real-time PCR assay utilizing a TaqMan MGB probe targeting the conserved 28S rDNA region. The assay demonstrated exceptional specificity, with no cross-reactivity against closely related fungal taxa or common co-occurring pathogens. The method exhibited high sensitivity, achieving a limit of detection (LOD) of 680 fg of genomic DNA. Furthermore, the assay was successfully validated using simulated environmental samples, where it accurately identified Colletotrichum within complex fungal communities. By providing a robust platform for genus-level screening, this methodology significantly enhances the efficiency of phytosanitary inspections and clinical diagnostics, facilitating timely biosecurity interventions and therapeutic decisions.

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