simplified automated image analysis for detection and phenotyping of mycobacterium tuberculosis on porous supports by monitoring growing microcolonies简化自动图像分析检测和表现型的结核分枝杆菌在多孔支持通过监测一个个独立王国.pdfVIP

Simplified Automated Image Analysis for Detection and Phenotyping of Mycobacterium tuberculosis on Porous Supports by Monitoring Growing Microcolonies 1 2 3 4 1 Alice L. den Hertog *, Dennis W. Visser , Colin J. Ingham , Frank H. A. G. Fey , Paul R. Klatser , Richard M. Anthony1 1 KIT Biomedical Research, Royal Tropical Institute, Amsterdam, The Netherlands, 2 Vleuten, The Netherlands, 3 Microdish BV, Houten, The Netherlands, 4 CCM Centre for Concepts in Mechatronics, Nuenen, The Netherlands Abstract Background: Even with the advent of nucleic acid (NA) amplification technologies the culture of mycobacteria for diagnostic and other applications remains of critical importance. Notably microscopic observed drug susceptibility testing (MODS), as opposed to traditional culture on solid media or automated liquid culture, has shown potential to both speed up and increase the provision of mycobacterial culture in high burden settings. Methods: Here we explore the growth of Mycobacterial tuberculosis microcolonies, imaged by automated digital microscopy, cultured on a porous aluminium oxide (PAO) supports. Repeated imaging during colony growth greatly simplifies ‘‘computer vision’’ and presumptive identification of microcolonies was achieved here using existing publically available algorithms. Our system thus allows the growth of individual microcolonies to be monitored and critically, also to change the media during the growth phase without disrupting the microcolonies. Transfer of identified microcolonies onto selective media allowed us, within 1-2 bacterial generations, to rapidly detect the drug susceptibility of individual