BMC veterinary researchJan 12, 2026

Fast and portable CRISPR test for detecting Mycoplasma gallisepticum infection in poultry

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Jun 27, 2026

Abstract

Essence

An mgc2-targeted RAA-CRISPR/Cas12a assay detected Mycoplasma gallisepticum within 1 hour using fluorescence or lateral-flow readouts.

Evidence

This diagnostic assay-development study optimized a poultry MG detection platform, found a 2 copies/uL detection limit and specificity against seven avian pathogens, and compared chicken samples with quantitative real-time PCR.

Caveat

The abstract does not report sample sizes, and clinical validation is described only as concordance with quantitative real-time PCR in chicken samples.

Simplified

BACKGROUND: Mycoplasma gallisepticum (MG) is a major pathogen that causes respiratory diseases 14in poultry, resulting in reduced production and severe economic losses. Current MG detection methods are time-consuming, labor-intensive, and expensive. Hence, the rapid and accurate detection of MG is critical for effective disease control. Therefore, this study aimed to develop a dual-mode diagnostic assay for sensitive and specific detection of MG by combining recombinase-aided amplification (RAA) with CRISPR/Cas12a technology. Conserved regions of the mgc2 gene were used for primer and CRISPR RNA design, and the reaction conditions were optimized to maximize detection efficiency.

RESULTS: The assay achieved a detection limit of 2 copies/µL and demonstrated high specificity against seven other common avian pathogens. Detection was visualized within 1 h using either fluorescence or lateral flow dipstick. Moreover, clinical validation of chicken samples showed complete concordance with quantitative real-time polymerase chain reaction results. Furthermore, an epidemiological investigation revealed that chickens had the highest positivity rate for MG among chickens, ducks, and pigeons in Hubei Province.

CONCLUSIONS: This simple, rapid, field-deployable method is valuable for timely MG surveillance and effective disease management in poultry production.