Hata Akihiko, Matsumori Katsuhito, Kitajima Masaaki, Katayama Hiroyuki
Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga, 520-0811, Japan,
Food Environ Virol. 2015 Mar;7(1):7-13. doi: 10.1007/s12560-014-9169-x. Epub 2014 Oct 18.
A viral adsorption-elution method using a flat/disk-type electronegative membrane (diameter of 47-90 mm) has been widely utilized to concentrate viruses in relatively small volumes of water (up to 10 L) due to limited filtration area. In the present study, we aimed to develop a virus concentration method that is based on the same principle and yet allows concentration of large volumes of water using a cartridge-type electronegative membrane. We modified two electronegative membrane-based methods for this purpose (i.e., Mg(2+) method and Al(3+) method) and determined recovery efficiencies of poliovirus and murine norovirus inoculated in water samples. The virus recovery efficiency of the Al(3+) method substantially decreased as the volume of water sample increased. In contrast, Mg(2+) method showed stable virus recovery efficiencies (10-54 %) even when 40 to 1,000 L of river and tap water samples were processed. The volume of the concentrate (400 mL) can be further reduced to 1.5 mL by a Centricon plus-70 centrifugal ultrafiltration device with overall recovery efficiencies of 8.8-16 %. Our results demonstrated that the newly developed virus concentration method enables detection of as low as 10(1) copies/L of viruses in water samples.
由于过滤面积有限，一种使用扁平/盘式电负性膜（直径47 - 90毫米）的病毒吸附 - 洗脱方法已被广泛用于在相对少量的水（最多10升）中浓缩病毒。在本研究中，我们旨在开发一种基于相同原理但能使用筒式电负性膜浓缩大量水的病毒浓缩方法。为此，我们对两种基于电负性膜的方法（即Mg(2+)法和Al(3+)法）进行了改进，并测定了接种于水样中的脊髓灰质炎病毒和鼠诺如病毒的回收效率。随着水样体积增加，Al(3+)法的病毒回收效率大幅下降。相比之下，即使处理40至1000升的河水和自来水样品，Mg(2+)法仍显示出稳定的病毒回收效率（10 - 54%）。通过Centricon plus - 70离心超滤装置可将浓缩物的体积（400毫升）进一步减少至1.5毫升，总体回收效率为8.8 - 16%。我们的结果表明，新开发的病毒浓缩方法能够检测出水样中低至10(1)拷贝/升的病毒。
