Detection of Campylobacter jejuni based on CRISPR/Cas12a with novel target yclQ obtained by pan-genome analysis

• Published in: Food science & technology 2024-05, Vol.199, p.116113, Article 116113
• Main Authors: Zhang, Weipei, Wang, Zhi, Ding, Yu, Huang, Zhenying, Zhao, Yibing, Pan, Qiqi, Xiao, Kaishan, Zhang, Jumei, Cai, Zhihe, Wu, Qingping, Wang, Juan
• Format: Article
• Language: English
• Subjects: Campylobacter jejuni; CRISPR/Cas12a; Novel detection target; PCR/RAA; yclQ
• ISSN: 0023-6438; 1096-1127
• DOI: 10.1016/j.lwt.2024.116113

Campylobacter jejuni (C. jejuni) is one of the most common causes of bacterial gastrointestinal foodborne infections worldwide. Conventional detection methods based on bacterial culture are time-consuming, therefore, rapid detection methods are urgently needed to be developed. The objective of this study was to screen a novel species-specific target and to develop a rapid and accurate method for the detection of C. jejuni in food. Through pan-genome analysis of 298 Campylobacter whole-genome sequence data, a new species-specific target yclQ was screened. The yclQ gene was verified through PCR to be unique to C. jejuni. Subsequently, based on the target yclQ, we developed of a CRISPR/Cas12a-based platform for fluorescence visualization and nucleic acid test strips. The RAA-CRISPR/Cas12a system demonstrates the ability to detect C. jejuni at 1.8 CFU/mL within 50 min. Compared to PCR (1.8 × 101 CFU/mL), the RAA and RAA-CRISPR/Cas12a assay showed higher sensitivity. The detection results of the actual samples were consistent with those of the isolation and culture procedures. In conclusion, the RAA-CRISPR/Cas12a system, based on the target yclQ, demonstrates significant potential for the rapid and accurate detection of C. jejuni in food. •A novel species-specific target yclQ for C. jejuni is obtained by pan-genome analysis.•A yclQ-based CRISPR/Cas12a-assisted nucleic acid detection platform is constructed.•The CRISPR/Cas12a platform based on yclQ can detect C. jejuni in food sample.