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Development of a Lateral Flow Strip-Based Recombinase-Aided Amplification for Active Chlamydia psittaci Infection
Jun Jiao1, Yong Qi2, Peisheng He1
1State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.
Abstract:
Chlamydia psittaci is the causative agent of psittacosis, a worldwide zoonotic disease. A rapid, specific, and sensitive diagnostic assay would be benefit for C. psittaci infection control. In this study, an assay combining recombinase-aided amplification and a lateral flow strip (RAA-LF) for the detection of active C. psittaci infection was developed. The RAA-LF assay targeted the CPSIT_RS02830 gene of C. psittaci and could be accomplished in 15 min at a single temperature (39°C). The analytical sensitivity of the assay was as low as 1 × 100 copies/μl and no cross-reaction with some other intracellular pathogens was observed. Moreover, all feces samples from mice infected with C. psittaci at day-1 post-infection were positive in the RAA-LF assay. In conclusion, the RAA-LF assay provides a convenient, rapid, specific and sensitive method for detection of active C. psittaci infection and it is also suitable for C. psittaci detection in field.
Insights
A new recombinase-aided amplification and lateral flow strip (RAA-LF) assay rapidly detects Chlamydia psittaci, the cause of psittacosis. This sensitive and specific diagnostic tool is ideal for field use in controlling this zoonotic disease.
Area of Science:
- Veterinary Microbiology
- Zoonotic Disease Research
- Molecular Diagnostics
Background:
- Chlamydia psittaci causes psittacosis, a significant global zoonotic disease.
- Effective control of C. psittaci infections necessitates rapid, specific, and sensitive diagnostic methods.
Purpose of the Study:
- To develop and validate a novel diagnostic assay for the detection of active Chlamydia psittaci infections.
- To create a tool that is both rapid and suitable for field applications.
Main Methods:
- Development of a recombinase-aided amplification and lateral flow strip (RAA-LF) assay.
- Targeting the CPSIT_RS02830 gene specific to C. psittaci.
- Assay validation using analytical sensitivity testing and cross-reactivity studies with other pathogens.
Main Results:
- The RAA-LF assay achieved detection as low as 1x10^0 copies/μl.
- The assay demonstrated high specificity, with no cross-reactivity observed against other tested intracellular pathogens.
- All fecal samples from mice infected with C. psittaci were positive at day-1 post-infection, indicating high sensitivity in vivo.
Conclusions:
- The developed RAA-LF assay offers a convenient, rapid, specific, and sensitive method for detecting active C. psittaci infections.
- The assay's performance characteristics make it suitable for on-site and field-based detection of C. psittaci.
- This diagnostic tool can aid in the timely control and management of psittacosis outbreaks.
