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Fast and reliable growth assessment is critical for validating in vitro drug susceptibility and high-throughput screening of newly developed drugs.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Modified media without yeast extract were evaluated for their ability to support the growth of <jats:italic>T. vaginalis<\/jats:italic> parasites. The potential of the nucleic acid-binding dye SYBR Green I for detecting <jats:italic>T. vaginalis<\/jats:italic> drug resistance was characterized, and seeding parasite concentration and incubation time were optimized. The fluorescence assay based on SYBR Green I was further validated in four <jats:italic>T. vaginalis<\/jats:italic> isolates with different susceptibilities to the antibiotics metronidazole, tinidazole, ornidazole and secnidazole, and compared with the traditional method that detects minimum lethal concentrations (MLCs).<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>A modified medium consisting of RPMI 1640 and Tryptone Plus as replacements for yeast extract and tryptone, respectively, in traditional trypticase-yeast extract-maltose (TYM) medium exhibited similar performance as TYM medium in maintaining <jats:italic>T. vaginalis<\/jats:italic> growth, while it showed much lower background fluorescent signals. The <jats:italic>T. vaginalis<\/jats:italic> SYBR Green I-based fluorescence (TSF) drug assay was found to have to satisfy one of two conditions to demonstrate the 50% inhibitory concentration of metronidazole for the sensitive isolate TV-334: (i) a seeding density of 3\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> parasites\/ml and an incubation time of 48\u00a0h; or (ii) a seeding density of 1\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> parasites\/ml and an incubation time of 72\u00a0h. Subsequent validation experiments revealed that the 48-h incubation\/3\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> parasites\/ml seeding density condition had a greater sensitivity to detect drug resistance than the 72-h condition. The TSF assay also exhibited high efficiency in identifying parasite drug resistance, as evidenced by its strong correlation with the standard MLC assay results (<jats:italic>P<\/jats:italic>\u2009=\u20090.003).<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>This study presents a robust TSF assay that has the potential to facilitate high-throughput, automated in vitro anti-trichomoniasis susceptibility testing for drug resistance monitoring and drug development. In comparison to the standard MLC method, this assay offers the advantages of reduced labor and elimination of subjective examination.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical Abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-023-05919-6","type":"journal-article","created":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T14:02:29Z","timestamp":1695045749000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A fluorescence-based assay for Trichomonas vaginalis drug screening"],"prefix":"10.1186","volume":"16","author":[{"given":"Qianqian","family":"Chen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingzhong","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhensheng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Meng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zenglei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2023,9,18]]},"reference":[{"key":"5919_CR1","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/j.idc.2023.02.001","volume":"37","author":"OT Van Gerwen","year":"2023","unstructured":"Van Gerwen OT, Opsteen SA, Graves KJ, Muzny CA. 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