State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, 130062, People's Republic of China.
Pingdu People's Hospital, Qingdao, 266700, People's Republic of China.
Parasitol Res. 2024 Apr 4;123(4):176. 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 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.
贾第虫病是由十二指肠贾第虫引起的一种常见肠道感染，给人类和家畜带来了重大的经济和健康负担。目前，迫切需要一种方便、有效的检测方法。由于其高效性和敏感性，基于 CRISPR/Cas12a 的诊断方法已被广泛应用于基于核酸的病原体检测。本研究建立了一种结合 CRISPR/Cas12a 和 RPA 的技术，可通过肉眼以高灵敏度（10 拷贝/μL）和特异性（与 9 种常见病原体无交叉反应）检测粪便样本中的 G. 十二指肠。在临床评估中，基于 RPA-CRISPR/Cas12a 的检测方法分别检测到 2%（1/50）的人粪便样本和 47%（33/70）的牛粪便样本中的贾第虫阳性，与巢式 PCR 的结果一致。我们的研究表明，用于检测 G. 十二指肠的 RPA-CRISPR/Cas12a 技术稳定、高效、灵敏、特异，且设备要求低。该技术为偏远和贫困地区的现场检测提供了新的机会。
