title: Sensitive and rapid detection of Chlamydia trachomatis by recombinase polymerase amplification directly from urine samples.
abstract: Chlamydia trachomatis is the most common sexually transmitted human pathogen. Infection results in minimal to no symptoms in approximately two-thirds of women and therefore often goes undiagnosed. C. trachomatis infections are a major public health concern because of the potential severe long-term consequences, including an increased risk of ectopic pregnancy, chronic pelvic pain, and infertility. To date, several point-of-care tests have been developed for C. trachomatis diagnostics. Although many of them are fast and specific, they lack the required sensitivity for large-scale application. We describe a rapid and sensitive form of detection directly from urine samples. The assay uses recombinase polymerase amplification and has a minimum detection limit of 5 to 12 pathogens per test. Furthermore, it enables detection within 20 minutes directly from urine samples without DNA purification before the amplification reaction. Initial analysis of the assay from clinical patient samples had a specificity of 100% (95% CI, 92%-100%) and a sensitivity of 83% (95% CI, 51%-97%). The whole procedure is fairly simple and does not require specific machinery, making it potentially applicable in point-of-care settings.
authors: Institute of Technology, University of Tartu, Tartu, Estonia. Electronic address: katrin.krolov@ut.ee.
authors: Katrin
authors: K
authors: Krõlov
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Jekaterina
authors: J
authors: Frolova
authors: Institute of Technology, University of Tartu, Tartu, Estonia; Department of Functional Genomics and Experimental Pathology Cluj-Napoca, I. Chiricuta Cancer Institute, Cluj-Napoca, Romania.
authors: Oana
authors: O
authors: Tudoran
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Julia
authors: J
authors: Suhorutsenko
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Taavi
authors: T
authors: Lehto
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Hiljar
authors: H
authors: Sibul
authors: Institute of Technology, University of Tartu, Tartu, Estonia.
authors: Imre
authors: I
authors: Mäger
authors: Tartu University Hospital's Women's Clinic and Tartu Sexual Health Clinique, Tartu, Estonia.
authors: Made
authors: M
authors: Laanpere
authors: Institute of Chemistry, University of Tartu, Tartu, Estonia; Selfdiagnostics OÜ, Tallinn, Estonia.
authors: Indrek
authors: I
authors: Tulp
authors: Institute of Technology, University of Tartu, Tartu, Estonia; Department of Neurochemistry, Stockholm University, Stockholm, Sweden.
authors: Ülo
authors: Ü
authors: Langel
mesh_terms: D002690:Chlamydia Infections / Q000175:diagnosis* / Q000235:genetics / Q000652:urine*; D002692:Chlamydia trachomatis / Q000201:enzymology / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry; D002798:Diacylglycerol Cholinephosphotransferase / Q000235:genetics; D005260:Female; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D045522:Recombinases / Q000737:chemistry; D012680:Sensitivity and Specificity; D015231:Sexually Transmitted Diseases, Bacterial / Q000175:diagnosis* / Q000235:genetics / Q000652:urine*
chemical_list: D004269:DNA, Bacterial; D045522:Recombinases; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D004259:DNA-Directed DNA Polymerase; D002798:Diacylglycerol Cholinephosphotransferase