Development of a virus concentration method and its application to detection of enterovirus and Norwalk virus from coastal seawater
Development of a virus concentration method and its application to detection of enterovirus and Norwalk virus from coastal seawater
复制标题
DOI：
10.1128/aem.68.3.1033-1039.2002
复制
发表时间：
2002-03-01
影响因子：
4.4
通讯作者：
Ohgaki, S
中科院分区：
文献类型：
--
作者：
Katayama, H;Shimasaki, A;Ohgaki, S
我们开发了一种新的程序，使用带负电荷的膜从水中浓缩肠道病毒。在用1 mM NaOH（pH 10.5）洗脱病毒之前，用0.5 mM H2SO 4（pH 3.0）冲洗膜以去除阳离子，分别促进了应用于纯水时脊髓灰质炎病毒回收率从33%提高到95%，应用于日本东京湾天然海水时脊髓灰质炎病毒回收率从38%提高到89%。该方法显示，从1升人工海水中加标脊髓灰质炎病毒的平均回收率为62%（n = 8）。当应用于海水时，该方法显示出比使用带正电荷的膜的常规方法（6%）更高的回收率（>61%）。该方法也不需要牛肉提取物洗脱，这在随后的通过逆转录-PCR进行的病毒基因组检测中具有抑制作用。用这种病毒浓缩方法，在冬季从2升东京湾水中检测到自然发生的诺瓦克病毒，在夏季从2升娱乐沿海海水中检测到传染性肠道病毒。
We developed a new procedure for concentration of enteric viruses from water using a negatively charged membrane. Rinsing the membrane with 0.5 mM H2SO4 (pH 3.0) in order to elute cations prior to viral elution with 1 mM NaOH (pH 10.5) promoted poliovirus recovery yields from 33 to 95% when applied to pure water and 38 to 89% when applied to natural seawater from Tokyo Bay, Japan, respectively. This method showed average recovery yields of spiked poliovirus of 62% (n = 8) from 1 liter of artificial seawater. This method showed higher recovery yields (>61%) than that of the conventional method using positively charged membrane (6%) when applied to seawater. This method is also free from beef extract elution, which has an inhibitory effect in the subsequent viral genome detection by reverse transcription-PCR. Naturally occurring Norwalk viruses from 2 liters of Tokyo Bay water in winter and infectious enteroviruses from 2 liters of recreational coastal seawater in summer were detected by using this viral concentration method.