Development and clinical validation of a loop-mediated isothermal amplification method for the rapid detection of Neisseria meningitidis

Loop-mediated isothermal amplification (LAMP) is an innovative technique that allows the rapid detection of target nucleic acid sequences under isothermal conditions without the need for complex instrumentation. The development, optimization, and clinical validation of a LAMP assay targeting the ctrA gene for the rapid detection of capsular Neisseria meningitidis were described. Highly specific detection of capsular N. meningitidis type strains and clinical isolates was demonstrated, with no cross-reactivity with other Neisseria spp. or with a comprehensive panel of other common human pathogens. The lower limit of detection was 6 ctrA gene copies detectable in 48 min, with positive reactions readily identifiable visually via a simple color change. Higher copy numbers could be detected in as little as 16 min. When applied to a total of 394 clinical specimens, the LAMP assay in comparison to a conventional TaqMan® based real-time polymerase chain reaction system demonstrated a sensitivity of 100% and a specificity of 98.9% with a ? coefficient of 0.942. The LAMP method represents a rapid, sensitive, and highly specific technique for the detection of N. meningitidis and has the potential to be used as a point-of-care molecular test and in resource-poor settings.

COHERENCE AND DOUBLE-PASS AMPLIFICATION IN GE SOFT-X-RAY LASER

From measurements of spatial coherence and beam divergence of Ge soft x-ray laser at a far field, the x-ray laser beam has been characterized as a partially coherent Gaussian beam. Double-pass amplification will improve spatial and temporal coherence, spectral brightness and efficiency. Close to 100% geometrical coupling efficiency has been obtained in double pass amplification in Ge. Transient loss of feedback is attributed to mirror structure damage within the build-up time of the x-ray laser. Prospect for generation of coherent x-ray laser beam is discussed.