Development of a MALDI-TOF nucleic acid mass spectrometry-based assay for the detection of 10 major avian diseases.

MALDI-TOF multiplex assay enables rapid, cost-effective simultaneous detection of 10 avian pathogens.

This study presents a novel MALDI-TOF mass spectrometry-based multiplex assay for simultaneous detection of 10 major avian pathogens, including three subtypes of avian influenza (H5/H7/H9), Newcastle disease, infectious bronchitis, infectious laryngotracheitis, fowl pox, avian adenovirus, and avian metapneumovirus. The method employs optimized multiplex PCR with specific primer design followed by SAP treatment and rapid mass spectrometry identification. The assay demonstrates excellent analytical performance with detection limits ranging from 4.70 to 52.54 copies/μL, exhibiting high sensitivity and specificity with reliable reproducibility across simulated samples. The technique significantly reduces detection time and cost compared to conventional diagnostic methods. This MALDI-TOF multiplex approach represents an important advancement in rapid diagnosis and surveillance of economically significant avian diseases, offering practical value for disease prevention and control in poultry operations.

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