Karim Q N, Logan R P, Puels J, Karnholz A, Worku M L
Department of Microbiology, Imperial College of Science, Technology and Medicine at St Mary's Hospital, London, UK.
J Clin Pathol. 1998 Aug;51(8):623-8. doi: 10.1136/jcp.51.8.623.
(1) To make precise measurements and comparisons of various aspects of motility of three gastrointestinal pathogens, Helicobacter pylori, Campylobacter jejuni, and Escherichia coli, in log phase growth; (2) to provide background information on motility data to study the influence of pH, viscosity, and chemotactic factors, thereby gaining a better understanding of bacterial pathogenesis.
Computer image processing technology and phase contrast microscopy (Hobson BacTracker) were used to measure several indices of bacterial motility in real time. Ten clinical isolates of each species in log phase liquid culture were studied.
C jejuni moved fastest, with a median curvilinear velocity (CLV) of 38.76 microns/s (range 29.08 to 52.82). Next was H pylori, median CLV 25.02 microns/s (range 12.07 to 29.07). E coli was the slowest, median CLV 12.73 microns/s (range 8.20 to 18.04). The straight line velocities showed similar trends. Measurement of track linearity (TL) showed that C jejuni moved the straightest (TL 60.3%), H pylori moved in wide circles (TL 28.7%), and E coli showed spinning movement without much linear displacement (TL 18.3%). There were significant differences in these three variables between the species studied, but no significant differences in measurements of time and frequency of halts between movement runs.
The BacTracker provides a useful technical aid for measuring several indices of bacterial motility objectively, reproducibly, and precisely, which is difficult to achieve without computer assistance. Accurate quantification of motility provides a basis for studying the factors which influence bacterial motility. It can provide phenotypic measurements of the effect of flagellar gene depletion.
(1)对处于对数生长期的三种胃肠道病原体幽门螺杆菌、空肠弯曲菌和大肠杆菌的运动各方面进行精确测量和比较;(2)提供运动数据的背景信息,以研究pH值、粘度和趋化因子的影响,从而更好地理解细菌发病机制。
使用计算机图像处理技术和相差显微镜(霍布森细菌追踪仪)实时测量细菌运动的几个指标。对每种处于对数期液体培养的10株临床分离株进行了研究。
空肠弯曲菌移动速度最快,曲线中位数速度(CLV)为38.76微米/秒(范围为29.08至52.82)。其次是幽门螺杆菌,CLV中位数为25.02微米/秒(范围为12.07至29.07)。大肠杆菌最慢,CLV中位数为12.73微米/秒(范围为8.20至18.04)。直线速度呈现相似趋势。轨迹线性度(TL)测量显示,空肠弯曲菌移动最直(TL为60.3%),幽门螺杆菌呈大圈移动(TL为28.7%),大肠杆菌表现为旋转运动且线性位移不大(TL为18.3%)。在所研究的物种之间,这三个变量存在显著差异,但运动过程中停顿的时间和频率测量无显著差异。
细菌追踪仪为客观、可重复且精确地测量细菌运动的几个指标提供了有用的技术辅助,若无计算机辅助则难以实现。运动的准确量化为研究影响细菌运动的因素提供了基础。它可以提供鞭毛基因缺失效应的表型测量。