Abstract
The global dissemination of the ceftriaxone-resistant Neisseria gonorrhoeae FC428 clone poses substantial challenges in the control and treatment of gonorrhoea, resulting in heightened morbidity rates and the increased risk of public health emergencies. However, currently, the identification of N. gonorrhoeae FC428 clone is labor-intensive and laboratory-based, which is not suitable for rapid on-site analyses and effective decision making. Herein, an Equipment-free CRISPR/Cas12a assisted RPA platform is developed for rapid duplex detection (E-card), which provides a solution for early and effective clinical diagnosis of N. gonorrhoeae, particularly the simultaneous tracking of ceftriaxone-resistant FC428 clone to allow reasonable and timely antimicrobial therapy. The E-card achieves 1 copy/reaction for N. gonorrhoeae screening and 10 copies/reaction for ceftriaxone-resistant FC428 clone detection within 40 min. The accuracy of N. gonorrhoeae and FC428 identification has been validated by concordance rates of 100 % and 98 % between the E-card platform and PCR-sequencing, respectively. This convenient CRISPR-based disposable molecular diagnostic platform can serve as a powerful tool to rapid detection of N. gonorrhoeae and its ceftriaxone-resistant FC428 clone, enabling effective assessment of infection, effective medicine decision making and the mitigation of health burdens.
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