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

被引:0
|
作者
Zhiteng Zhao
Songgao Cao
Min Sun
Qiankun Yang
Taojun Huang
Xing Yang
Jianhua Li
Xichen Zhang
Xin Li
Xiaocen Wang
Weina Jiang
Pengtao Gong
机构
[1] Jilin University,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
[2] Pingdu People’s Hospital,Integrated Laboratory of Pathogenic Biology, College of Preclinical Medicine
[3] Dali University,Department of Pathology
[4] Qingdao Municipal Hospital,undefined
来源
Parasitology Research | 2024年 / 123卷
关键词
RPA-CRISPR/Cas12a; Visual detection; Recombinase polymerase amplification; Lateral flow strips;
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摘要
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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