TtrAgo-mediated nucleic acid detection system and portable device for rapid detection of sexually transmitted diseases

The development of rapid and multiplexed point-of-care (POC) diagnostic tools is vital for the prevention and control of sexually transmitted diseases (STIs). Here, we developed a POC-comprehensive Thermococcus thioreducensArgonaute (TtrAgo)-mediated nucleic acid detection system (POC-CANDY) and palm-sized portable detection device "Owl-1" for the simultaneous detection of Ureaplasma urealyticum, Chlamydia trachomatis, Neisseria gonorrhoeae, human papillomavirus types 16/18 and antibiotic resistance molecular markers [tetM, and gyrA mutation (S91F)]. Using recombinase polymerase amplification (RPA), the optimized POC-CANDY could finish the whole detection procedure within 55 min and achieve a limit of detection of 10 copies/μL. When validated by clinical STI samples, POC-CANDY showed 100% consistency with quantitative PCR. Additionally, compared with the PfAgo-based system, POC-CANDY significantly improved the sensitivity of distinguishing single nucleotide variations. The results demonstrated that POC-CANDY can be easily applied locally or on site. This study also promotes the utility of the TtrAgo-mediated technique in clinical diagnosis.