Rapid visual detection of Giardia duodenalis in faecal samples using an RPA-CRISPR/Cas12a system

• Published in: Parasitology research (1987) Vol. 123; no. 4; p. 176
• Main Authors: Zhao, Zhiteng, Cao, Songgao, Sun, Min, Yang, Qiankun, Huang, Taojun, Yang, Xing, Li, Jianhua, Zhang, Xichen, Li, Xin, Wang, Xiaocen, Jiang, Weina, Gong, Pengtao
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.04.2024; Springer Nature B.V
• Subjects: Animals; Biological Assay; Biomedical and Life Sciences; Biomedicine; Cattle; CRISPR; CRISPR-Cas Systems; Cross-reactivity; Feces; Giardia - genetics; Giardia duodenalis; Giardia lamblia - genetics; Giardiasis; Giardiasis - diagnosis; Giardiasis - veterinary; Humans; Immunology; Livestock; Medical Microbiology; Microbiology; Pathogens; RPA-CRISPR/Cas12a; Visual detection; Lateral flow strips; Recombinase polymerase amplification; Giardia duodenalis
• ISSN: 0932-0113; 1432-1955
• DOI: 10.1007/s00436-024-08197-y

Giardiasis is a common intestinal infection caused by Giardia duodenalis , which is a major economic and health burden for humans and livestock. Currently, a convenient and effective detection method is urgently needed. CRISPR/Cas12a-based diagnostic methods have been widely used for nucleic acid-based detection of pathogens due to their high efficiency and sensitivity. In this study, a technique combining CRISPR/Cas12a and RPA was established that allows the detection of G. duodenalis in faecal samples by the naked eye with high sensitivity (10 −1 copies/μL) and specificity (no cross-reactivity with nine common pathogens). In clinical evaluations, the RPA-CRISPR/Cas12a-based detection assay detected Giardia positivity in 2% (1/50) of human faecal samples and 47% (33/70) of cattle faecal samples, respectively, which was consistent with the results of nested PCR. Our study demonstrated that the RPA-CRISPR/Cas12a technique for G. duodenalis is stable, efficient, sensitive, specific and has low equipment requirements. This technique offers new opportunities for on-site detection in remote and poor areas.