{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:48:43Z","timestamp":1792788523602,"version":"4.5.0"},"update-to":[{"DOI":"10.1371\/journal.pntd.0012753","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T00:00:00Z","timestamp":1735171200000}}],"reference-count":39,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T00:00:00Z","timestamp":1733961600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Technology Research and Development Program of Sichuan Province","award":["2022YFN0013"],"award-info":[{"award-number":["2022YFN0013"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"\n Background<\/jats:title>\n \n Cystic echinococcosis (CE), caused by\n Echinococcus granulosus<\/jats:italic>\n sensu lato (\n E<\/jats:italic>\n .\n granulosus<\/jats:italic>\n s.l.), remains a significant zoonotic parasitic disease affecting both livestock and humans. It arises from the ingestion of food and water contaminated with canine feces containing\n E<\/jats:italic>\n .\n granulosus<\/jats:italic>\n eggs. The detection of these eggs in canine feces is essential for guiding effective preventative measures against the disease. Therefore, the development of a novel accurate, rapid, and visually interpretable point-of-care test is crucial for controlling CE.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n We combined recombinase polymerase amplification (RPA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) with a CRISPR-associated protein 12a (Cas12a) system, forming the RPA-CRISPR\/Cas12a assay. This assay targeted the\n E<\/jats:italic>\n .\n granulosus<\/jats:italic>\n mitochondrial\n nad2<\/jats:italic>\n gene and utilized a lateral flow strip for visual readout. To improve field applicability, we integrated a simple and cost-effective NaOH-Based DNA extraction method. Clinical validation included testing DNA extracted from eighteen canine fecal samples, followed by comparison with quantitative PCR (qPCR) and two commercial enzyme-linked immunosorbent assay (ELISA) kits.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The RPA-CRISPR\/Cas12a assay showed a detection limit of 1 fg\/\u03bcL DNA, without any cross-reactivity with related tapeworms such as\n Echinococcus multilocularis<\/jats:italic>\n ,\n Dipylidium caninum<\/jats:italic>\n ,\n Taenia hydatigera<\/jats:italic>\n ,\n Taenia multiceps<\/jats:italic>\n , and\n Taenia pisiformis<\/jats:italic>\n . When applied to 62 clinical fecal samples from dogs, the RPA-CRISPR\/Cas12a assay demonstrated 68% sensitivity, while the developed RPA-CRISPR\/Cas12a-NaOH assay exhibited 45% sensitivity. In the field performance comparison of the RPA-CRISPR\/Cas12a and the RPA-CRISPR\/Cas12a-NaOH assay with qPCR and two ELISA kits, the sensitivity, consistency rate, and Youden\u2019s index suggested good or fair agreement with the currently employed detection methods.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n This study describes the development and validation of the RPA-CRISPR\/Cas12a and RPA-CRISPR\/Cas12a-NaOH assays for detecting\n E<\/jats:italic>\n .\n granulosus<\/jats:italic>\n in canine feces. The developed assays surpassed previous detection methods in providing enhanced diagnostic sensitivity and enabling point-of-care testing. Moreover, these assays hold potential for surveilling\n E<\/jats:italic>\n .\n granulosus<\/jats:italic>\n in low-income countries.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1371\/journal.pntd.0012753","type":"journal-article","created":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T13:49:07Z","timestamp":1734011347000},"page":"e0012753","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["Development of a Copro-RPA-CRISPR\/Cas12a assay to detect Echinococcus granulosus nucleic acids isolated from canine feces using NaOH-based DNA extraction 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