title: Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample.
abstract: Chlamydia trachomatis is an obligate intracellular human pathogen and is the most common cause of sexually transmitted diseases affecting both men and women. The pathogen can cause prostatitis and epididymitis in men. In women, cervicitis, pelvic inflammatory disease, ectopic pregnancy and acute or chronic pelvic pain are frequent complications. More than half of C. trachomatis-positive patients have minimal or no symptoms, providing an ongoing reservoir for the infection. The lack of sensitive large-scale applicable point- of- care (POC) tests for C. trachomatis detection makes it difficult to diagnose chlamydia infection efficiently in resource-limited environments.
  
  
  
  A rapid and sensitive assay based on loop-mediated isothermal amplification method (LAMP) was combined with antimicrobial peptide lysis, which is able to detect at least 7 C. trachomatis pathogens per reaction directly from urine samples.
  
  
  
  Our study comprising 91 first-void urine samples showed that specificity of the assay is 100 % and sensitivity 73 % when using antimicrobial peptide lysis mix. Additionally we demonstrate that our assay does not give any cross-reactivity with 30 pathogen's DNA potentially present in the urine samples. Furthermore, the assay's novel approach does not require purification or extraction of DNA from clinical sample prior to amplification, so the need for specialized equipment is eliminated.
  
  
  
  The whole procedure is significantly less laborious, less time-consuming and consequently less expensive for early detection and identification of infectious disease. C. trachomatis specific LAMP assay is relatively simple to perform and could therefore be applied in numerous POC settings.
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia. iriss@ut.ee.
authors.forename: Jekaterina
authors.initials: J
authors.lastname: Jevtuševskaja
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia.|SelfD Technologie GmbH, Leipzig, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Uusna
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia.
authors.forename: Liis
authors.initials: L
authors.lastname: Andresen
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia.
authors.forename: Katrin
authors.initials: K
authors.lastname: Krõlov
authors.affiliation: The Tartu University Hospital's Women's Clinic and Tartu Sexual Health Clinic, Tartu, Estonia.
authors.forename: Made
authors.initials: M
authors.lastname: Laanpere
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia.
authors.forename: Tiia
authors.initials: T
authors.lastname: Grellier
authors.affiliation: SelfD Technologie GmbH, Leipzig, Germany.|University of Tartu, Institute of Chemistry, Tartu, Estonia.
authors.forename: Indrek
authors.initials: I
authors.lastname: Tulp
authors.affiliation: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu, 50411, Estonia.|Stockholm University, Stockholm, Sweden.
authors.forename: Ülo
authors.initials: Ü
authors.lastname: Langel
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D002690:Chlamydia Infections / Q000175:diagnosis*; D002692:Chlamydia trachomatis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D011247:Pregnancy; D012680:Sensitivity and Specificity
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D004269:DNA, Bacterial
keywords: Chlamydia trachomatis; Diagnostics of sexually transmitted diseases; Loop-mediated isothermal amplification; Pathogen detection in crude urine; Point-of-care assay