{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T12:22:56Z","timestamp":1768479776796,"version":"3.49.0"},"reference-count":52,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2023,1,22]],"date-time":"2023-01-22T00:00:00Z","timestamp":1674345600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81802028"],"award-info":[{"award-number":["81802028"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Eukaryotic Microbiology"],"published-print":{"date-parts":[[2023,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Trichomoniasis is a common and curable sexually transmitted disease worldwide. The rapid, convenient, and accurate diagnosis of trichomoniasis is an important link in the prevention and treatment of the disease. The current detection methods of <jats:italic>Trichomonas vaginalis<\/jats:italic> are mainly wet mount microscopy, culture, nested PCR, and loop\u2010mediated isothermal amplification. However, these detection methods have some shortcomings. In this study, a recombinant enzyme polymerase amplification (RPA) assay had been conducted to detect <jats:italic>T. vaginalis<\/jats:italic>. The target gene and the corresponding primers were screened, and the reaction system and conditions were optimized in the assay of RPA. The sensitivity and specificity of this detection method were analyzed. The detection efficiency of wet mount microscopy, culture, nested PCR, and RPA was compared by testing 53 clinical samples from vaginal secretions. By screening, the actin gene of <jats:italic>T. vaginalis<\/jats:italic> could be used as a target gene for RPA detection of <jats:italic>T. vaginalis<\/jats:italic>, and the optimum reaction condition to amplify the actin gene by RPA was at 39\u00b0C for 30\u2009min. The detection limit of <jats:italic>T. vaginalis DNA<\/jats:italic> using RPA was 1\u00a0pg, corresponding to a sensitivity of approximately five trophozoites. The RPA assay demonstrated high specificity for <jats:italic>T. vaginalis<\/jats:italic>, and there was no cross\u2010reactivity with <jats:italic>Giardia lamblia<\/jats:italic>, <jats:italic>Escherichia coli<\/jats:italic>, <jats:italic>Lactobacillus<\/jats:italic>, <jats:italic>Toxoplasma gondii<\/jats:italic>, <jats:italic>Staphylococcus aureus<\/jats:italic>, and <jats:italic>Candida albicans<\/jats:italic>. Of the 53 clinical samples, the positive rates of <jats:italic>T. vaginalis<\/jats:italic> detected by wet mount microscopy, culture, nested PCR and RPA were 50.9 4% (27\/53), 71.7% (38\/53), 71.7% (38\/53), and 69.81% (37\/53), respectively. Compared with culture which was used as the gold standard for diagnosing trichomoniasis, testing clinical samples by wet mount microscopy showed 71.05% sensitivity, 100% specificity, and moderate diagnostic agreement with the culture (<jats:italic>K<\/jats:italic>\u00a0=\u00a00.581, <jats:italic>Z<\/jats:italic>\u00a0=\u00a04.661, <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001). The nested PCR showed 100% sensitivity, 100% specificity, and excellent diagnostic agreement (<jats:italic>K<\/jats:italic>\u00a0=\u00a01, <jats:italic>Z<\/jats:italic>\u00a0=\u00a07.28, <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001), while RPA displayed 97.37% sensitivity, 100% specificity, and excellent diagnostic agreement (<jats:italic>K<\/jats:italic>\u00a0=\u00a00.954, Z\u00a0=\u00a06.956, <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001). At the present study, rapid amplification of actin gene by RPA could be used as a tool for detection of <jats:italic>T. vaginalis<\/jats:italic>. The detection method of RPA was more sensitive than wet mount microscopy and displayed excellent specificity. Moreover, RPA amplification of actin gene did not require a PCR instrument and the amplification time was shorter than that of ordinary PCR. Therefore, the RPA assay was proposed in this study as a point\u2010of\u2010care examination and a diagnostic method of <jats:italic>T. vaginalis<\/jats:italic> infection, which exhibited the potential value in the treatment and prevention of trichomoniasis.<\/jats:p>","DOI":"10.1111\/jeu.12963","type":"journal-article","created":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T07:49:10Z","timestamp":1673509750000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Construction a novel detection method for <i>Trichomonas vaginalis<\/i> based on recombinant enzyme polymerase amplification targeting the Actin gene"],"prefix":"10.1111","volume":"70","author":[{"given":"Fakun","family":"Li","sequence":"first","affiliation":[{"name":"Xinxiang Key Laboratory of Pathogenic Biology, Department of Pathogenic Biology, School of Basic Medical Sciences Xinxiang Medical University  Xinxiang 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