title: Bench-scale experiments for the development of a unified loop-mediated isothermal amplification (LAMP) assay for the in vitro diagnosis of Leishmania species' promastigotes.
abstract: We developed, in bench-scale experiments, a unified loop-mediated isothermal amplification (LAMP) assay for the detection of cutaneous, mucocutaneous and visceral leishmaniasis using DNA of cultivated promastigotes. Two primer sets for the LAMP assay were designed based on the 18S rRNA gene, and their sensitivity and specificity were tested and compared. Both of them were specific for Leishmania as the DNA of all ten Leishmania species tested was amplified, whereas the DNA of other parasites, including that of Trypanosoma, was not. The detection limit for primer set 1 ranged between 30 pg and 3·6 fg, depending on which Leishmania species tested. Primer set 2 showed high sensitivity, but was less sensitive than primer set 1. Our findings lead to the conclusion that the LAMP assay with primer set 1 is a promising and effective assay for the successful detection of a wide range of Leishmania infections using only a unified multiplex LAMP test.
authors.affiliation: Medical and Molecular Parasitology Laboratory,University of Cologne,Medical School, Centre of Anatomy, Institute II, Cologne,Germany.
authors.forename: M
authors.initials: M
authors.lastname: Karani
authors.affiliation: Medical and Molecular Parasitology Laboratory,University of Cologne,Medical School, Centre of Anatomy, Institute II, Cologne,Germany.
authors.forename: I
authors.initials: I
authors.lastname: Sotiriadou
authors.affiliation: National Institute of Environmental Health,Department of Water Hygiene, Gyáli ut 2-6, Budapest,Hungary.
authors.forename: J
authors.initials: J
authors.lastname: Plutzer
authors.affiliation: Medical and Molecular Parasitology Laboratory,University of Cologne,Medical School, Centre of Anatomy, Institute II, Cologne,Germany.
authors.forename: P
authors.initials: P
authors.lastname: Karanis
mesh_terms: D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000175:diagnosis* / Q000469:parasitology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S