A recombinase polymerase amplification-based assay for rapid detection of Chlamydia psittaci

ABSTRACT Chlamydophila psittaci (C. psittaci) is a zoonotic agent of systemic wasting disease in birds, and atypical pneumonia in mammalians including humans, constituting a public health risk. A rapid diagnostic assay would be beneficial in screening C. psittaci in the field. In this study, we developed a probe-based recombinase polymerase amplification (RPA) assay for the rapid detection of C. psittaci. The specific primer pairs and probe targeting the conserved region of ompA gene were designed and applied to the real-time RT-RPA assay. The test can be performed at 39 °C for 20 min using a portable device, with sensitivities approaching 100 copies of DNA molecules per reaction, with no cross-reaction with other pathogens. The clinical performance of the RPA assay was evaluated in an outbreak of C. psittaci, and high accuracy levels in field applications. The epidemic C. psittaci strains were classed into two genotypes: A and C. Collectively, this study offers a promising approach in screening for C. psittaci both in a laboratory and in field settings, and can be used as an effective clinical test to monitor outbreaks in domestic fowl populations.