Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample

• Published in: BMC infectious diseases Vol. 16; no. 1; p. 329
• Main Authors: Jevtuševskaja, Jekaterina, Uusna, Julia, Andresen, Liis, Krõlov, Katrin, Laanpere, Made, Grellier, Tiia, Tulp, Indrek, Langel, Ülo
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 13.07.2016; BioMed Central
• Subjects: Analysis; Anti-Infective Agents - pharmacology; Antimicrobial Cationic Peptides - pharmacology; Antimicrobial peptides; Chlamydia infections; Chlamydia Infections - diagnosis; Chlamydia trachomatis; Chlamydia trachomatis - drug effects; Chlamydia trachomatis - genetics; Chlamydia trachomatis - isolation & purification; Complications and side effects; Diagnosis; Diagnostics of sexually transmitted diseases; DNA, Bacterial - isolation & purification; DNA, Bacterial - metabolism; DNA, Bacterial - urine; Female; Humans; Infectious diseases; Limit of Detection; Loop-mediated isothermal amplification; Male; Nucleic Acid Amplification Techniques - methods; Pathogen detection in crude urine; Point-of-care assay; Point-of-Care Systems; Pregnancy; Risk factors; Sensitivity and Specificity; Sexually transmitted diseases; Diagnostics of sexually transmitted diseases; Loop-mediated isothermal amplification; Chlamydia trachomatis; Pathogen detection in crude urine; Point-of-care assay
• ISSN: 1471-2334; 1471-2334
• DOI: 10.1186/s12879-016-1674-0

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.