Abstract
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.
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No datasets were generated or analysed during the current study.
Abbreviations
- LFS:
-
Lateral flow strips
- RPA:
-
Recombinase polymerase amplification
- CRISPR:
-
Clustered regularly interspaced short palindromic repeats
- Cas:
-
CRISPR-associated proteins
- crRNA:
-
CRISPR RNA
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Funding
This work was funded by the China Wool-sheep & Cashmere-goat Research System (Grant No. CARS-39), the Jilin Scientific and Technological Development Program (Grant No. 20220508101RC, 20240404011ZP), and the Changchun Science and Technology Program (Grant No. 21ZY24).
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ZTZ and MS designed the study. XL, TJH, and SGC collected data. ZTZ, MS, XCW and XY analyzed data. ZTZ and QKY interpreted data and wrote the manuscript. PTG, WNJ, JHL and XCZ revised the manuscript from preliminary draft to submission. All authors contributed to the article. All authors read and approved the final manuscript.
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The study involving human participants was reviewed and approved by the Medical Ethics Committee of the Second Hospital of Jilin University, Changchun, China, 2022 Research and Review No. (118).
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Zhao, Z., Cao, S., Sun, M. et al. Rapid visual detection of Giardia duodenalis in faecal samples using an RPA-CRISPR/Cas12a system. Parasitol Res 123, 176 (2024). https://doi.org/10.1007/s00436-024-08197-y
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DOI: https://doi.org/10.1007/s00436-024-08197-y