Application of Pyrococcus furiosus Argonaute coupled with RPA for highly sensitive and specific detection of Mycoplasma gallisepticum.

Abstract

Mycoplasma gallisepticum (MG) is the pathogen that causes chronic respiratory disease (CRD) in chickens and infectious sinusitis in turkeys. Clinical symptoms such as cough and dyspnea cause a decrease in the egg production rate and carcass quality of chickens, which results in considerable economic losses to the poultry industry. Thus, it is urgent to establish a fast, accurate and sensitive method for detecting MG. In this study, the detection of MG by combining recombinase polymerase amplification (RPA) with argonaute from Pyrococcus furiosus (PfAgo) was established that allows visual observation under blue and UV light. The sensitivity of RPA-PfAgo reached 1 copy/µL. After the chicken infection model, it was found that tracheal inhalation was superior to eye and nose drips and sprays, the results of RPA-PfAgo and qPCR were completely consistent. This study provides a reference tool for the monitoring of MG in the clinic and contributes to the early diagnosis of the disease.