Beniac Daniel R, Siemens Christine G, Wright Christine J, Booth Tim F
Viral Diseases Division, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB R3E 3P6, Canada.
Viruses. 2014 Sep 19;6(9):3458-71. doi: 10.3390/v6093458.
Diagnostic electron microscopy for infectious diseases has the advantage that "everything" in the specimen can be observed, without a priori knowledge of the likely identity of the microorganisms present in the sample. The classical specimen preparation method used employs a droplet of sample, which allows particles to adsorb to a support film, and is subsequently negative stained. This "grid on drop" procedure has a sensitivity range of approximately 106 viruses per mL if no enrichment procedures are used. In the current investigation we present a novel use of filtration that allows us to detect viruses at concentrations as low as 102 viruses per mL. We present here methods based on filtration, in which total virus, and not virus concentration, is the limiting factor for detection. We show that filtration is more sensitive than conventional negative staining and can detect as few as 5 × 103 particles per sample.
可以观察样本中的“所有东西”，而无需预先了解样本中可能存在的微生物的身份。所采用的经典样本制备方法是使用一滴样本，使颗粒吸附到支撑膜上，随后进行负染色。如果不使用富集程序，这种“液滴上的网格”程序的灵敏度范围约为每毫升106个病毒。在当前的研究中，我们展示了过滤的一种新用途，它使我们能够检测低至每毫升102个病毒浓度的病毒。我们在此介绍基于过滤的方法，其中总病毒量而非病毒浓度是检测的限制因素。我们表明，过滤比传统的负染色更灵敏，每个样本可检测低至5×103个颗粒。
