Evaluation of nucleic acid isothermal amplification methods for human clinical microbial infection detection

F. Urban
Spartan Innovations, Michigan, United States

Keywords: infectious disease, diagnostic, infection, detection, pathogens

Identification of infection with rapid antimicrobial treatment are primary goals of medical care, but precise identification of offending organisms is slow and broad spectrum empirical therapy is employed to cover most potential pathogens. Current methods for identification of bacterial pathogens in a clinical setting typically require days of time, or a four to eight hour growth phase followed by DNA extraction, purification and nucleic acid based amplification. We demonstrate rapid genetic diagnostics methods utilizing loop-mediated isothermal amplification (LAMP) to test for 15 common infection pathogen targets (In-Dx). The method, utilizing filtration to rapidly concentrate bacteria in sample matrices with lower bacterial loads and direct LAMP amplification from clinical blood, urine, wound, sputum and stool samples, takes 70-120 minutes to final results. The panel was tested using two methods of detection: 1) real-time LAMP amplification and 2) visual discrimination of color change following amplification. Duplicate clinical blood, urine, wound/swab, sputum and stool samples collected from prospectively-enrolled patients with suspected infection were analyzed by LAMP and compared to hospital testing for detecting targeted clinical pathogens across culture types. Results indicate the LAMP-based panel allows rapid and precise diagnosis of clinical infections by targeted pathogens across multiple culture types for point-of-care utilization.