title: Development of a loop-mediated isothermal amplification assay for detection of Trichomonas vaginalis.
abstract: A loop-mediated isothermal amplification (LAMP) assay targeting the 2-kbp repeated DNA species-specific sequence was developed for detection of Trichomonas vaginalis, the causative agent of trichomoniasis. The analytical sensitivity and specificity of the LAMP assay were evaluated using pooled genital swab and urine specimens, respectively, spiked with T. vaginalis trophozoites. Genital secretion and urine did not inhibit the detection of the parasite. The sensitivity of the LAMP was 10-1000 times higher than the PCR performed. The detection limit of LAMP was 1 trichomonad for both spiked genital swab and urine specimens. Also, LAMP did not exhibit cross-reactivity with closely-related trichomonads, Trichomonas tenax and Pentatrichomonas hominis, and other enteric and urogenital microorganisms, Entamoeba histolytica, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. This is the first report of a LAMP assay for the detection of T. vaginalis and has prospective application for rapid diagnosis and control of trichomoniasis.
authors.affiliation: College of Medicine, University of the Philippines, Ermita, Manila 1000, Philippines.
authors.forename: John Carlo B
authors.initials: JC
authors.lastname: Reyes
authors.affiliation: Department of Parasitology, College of Public Health, University of the Philippines, Ermita, Manila 1000, Philippines.
authors.forename: Juan Antonio A
authors.initials: JA
authors.lastname: Solon
authors.affiliation: Institute of Biology, College of Science, University of the Philippines, Diliman, Quezon City 1101, Philippines; Molecular Protozoology Laboratory, Natural Sciences Research Institute, University of the Philippines, Diliman, Quezon City 1101, Philippines. Electronic address: wlrivera@science.upd.edu.ph.
authors.forename: Windell L
authors.initials: WL
authors.lastname: Rivera
mesh_terms: D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D014245:Trichomonas Infections / Q000175:diagnosis* / Q000469:parasitology; D014246:Trichomonas vaginalis / Q000235:genetics / Q000302:isolation & purification*
keywords: Analytical sensitivity and specificity; Diagnosis; LAMP; Trichomonas vaginalis