{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T21:07:54Z","timestamp":1788210474743,"version":"build-2803163510"},"reference-count":27,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T00:00:00Z","timestamp":1765497600000},"content-version":"vor","delay-in-days":345,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,1,1]],"date-time":"2025-01-01T00:00:00Z","timestamp":1735689600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32202839"],"award-info":[{"award-number":["32202839"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004479","name":"Natural Science Foundation of Jiangxi Province","doi-asserted-by":"publisher","award":["20242BAB20312"],"award-info":[{"award-number":["20242BAB20312"]}],"id":[{"id":"10.13039\/501100004479","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transboundary and Emerging Diseases"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:p>\n                    <jats:italic>Pentatrichomonas hominis (P. hominis)<\/jats:italic>\n                    and\n                    <jats:italic>Tritrichomonas foetus (T. foetus)<\/jats:italic>\n                    are prevalent intestinal protozoa.\n                    <jats:italic>P. hominis<\/jats:italic>\n                    is associated with chronic diarrhea in humans and animals, whereas\n                    <jats:italic>T. foetus<\/jats:italic>\n                    causes gastrointestinal disease in companion animals and reproductive\u2010tract infection in cattle. Rapid and accurate identification of these infections at the point\u2010of\u2010care (POC) is crucial for the diagnosis and effective management of zoonotic diseases. In this study, we developed two novel recombinase polymerase amplification (RPA) assays coupled with CRISPR\/Cas12a detection. The dual\u2010species assay, using a lateral\u2010flow format, targeted species\u2010specific regions of the 18S rRNA gene of\n                    <jats:italic>P. hominis<\/jats:italic>\n                    and\n                    <jats:italic>T. foetus<\/jats:italic>\n                    , and under ideal conditions, delivered visual results within 40\u2009min for a single sample at 37\u00b0C.\n                    <jats:italic>P. hominis<\/jats:italic>\n                    \u2010specific assay: To differentiate\n                    <jats:italic>P. hominis<\/jats:italic>\n                    in mixed infections with\n                    <jats:italic>T. foetus<\/jats:italic>\n                    , a second assay targeted the highly conserved\n                    <jats:italic>Spo11-1<\/jats:italic>\n                    gene of\n                    <jats:italic>P. hominis<\/jats:italic>\n                    . Optimal crRNA\u2010412 and RPA primers were selected for maximal Cas12a cleavage efficiency. Analytical sensitivity and specificity were compared with conventional nested polymerase chain reaction (PCR) and Sanger sequencing. The results showed that The dual\u2010species assay detected as few as 50 DNA copies\/\u00b5L of either parasite with no cross\u2010reactivity to\n                    <jats:italic>Giardia lamblia<\/jats:italic>\n                    ,\n                    <jats:italic>Cystoisospora canis<\/jats:italic>\n                    ,\n                    <jats:italic>Cryptosporidium<\/jats:italic>\n                    spp.,\n                    <jats:italic>Toxoplasma gondii<\/jats:italic>\n                    ,\n                    <jats:italic>Toxocara canis<\/jats:italic>\n                    , and\n                    <jats:italic>Toxascaris leonina<\/jats:italic>\n                    . Among 70 fecal samples of companion animal (48 dogs and 22 cats), 14 (29.2%) dogs tested positive for\n                    <jats:italic>P. hominis<\/jats:italic>\n                    , and eight cats (36.4%) tested positive for\n                    <jats:italic>T. foetus<\/jats:italic>\n                    by nested PCR. Due to financial and logistical constraints, we selected a smaller subset for subsequent analysis with the RPA\u2010CRISPR\/Cas12a lateral\u2010flow strip (LFS) assay, which showed 100% diagnostic concordance with PCR. The Spo11\u20101 assay achieved a limit of detection of 20 DNA copies\/\u00b5L and specifically recognized\n                    <jats:italic>P. hominis<\/jats:italic>\n                    among a panel that included seven non\u2010target protozoa and helminths. Validation on 10 additional canine and feline samples (four positives and six negatives) showed complete agreement with nested\u2010PCR results. In conclusion, this CRISPR\u2013based diagnostic approach significantly enhances the efficiency and accuracy of Trichomonads detection, offering a practical, cost\u2010effective solution particularly suitable for veterinary and potentially human healthcare diagnostics in resource\u2010limited settings.\n                  <\/jats:p>","DOI":"10.1155\/tbed\/9995679","type":"journal-article","created":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T07:23:36Z","timestamp":1765610616000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Emerging Trichomonad Infections in Companion Animals: Rapid Visual Detection of\n                    <i>Pentatrichomonas hominis<\/i>\n                    and\n                    <i>Tritrichomonas foetus<\/i>\n                    Using an RPA\u2010CRISPR\/Cas12a Assay"],"prefix":"10.1155","volume":"2025","author":[{"given":"Yang","family":"Zou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi-Wen","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying-Rui","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liu-Min","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5181-3728","authenticated-orcid":false,"given":"Xiao-Qing","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,12,12]]},"reference":[{"key":"e_1_2_14_1_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2025.1522176"},{"key":"e_1_2_14_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.1968.tb02151.x"},{"key":"e_1_2_14_3_2","doi-asserted-by":"publisher","DOI":"10.1177\/2055116918774959"},{"key":"e_1_2_14_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00436-019-06550-0"},{"key":"e_1_2_14_5_2","first-page":"602","article-title":"Survey and Molecular Characterization of Trichomonads in Pigs in Anhui Province, East China, 2014","volume":"13","author":"Li W. 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