Laboratory for Zoonoses and Environmental Microbiology, Centre for Infectious Disease Control Netherlands, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
J Appl Microbiol. 2009 Jul;107(1):97-105. doi: 10.1111/j.1365-2672.2009.04184.x. Epub 2009 Mar 3.
To assess public health risks of rotavirus via drinking water consumption, a cell culture-PCR assay was developed and optimized for the detection of infectious environmental rotavirus strains in naturally contaminated source waters for drinking water production.
Infectious rotavirus concentrations were estimated by an optimized cell culture-PCR assay as most probable numbers by using the presence or absence of replicated virus in different sample volumes. Infectious rotavirus was detected in 11 of 12 source water samples in concentrations varying from 0.19 (0.01-0.87) to 8.3 (1.8-34.0) infectious PCR detectable units per litre (IPDU/l), which was not significantly different from the concentrations of infectious enterovirus in these samples.
In 55% of the samples, rotavirus genomes were 1000 to 10 000 times (3 log(10)-4 log(10)) more abundantly present than infectious rotavirus particles, whereas in the remaining 45% of the samples, rotavirus genomes were less than 1000 times (<3 log(10)) more abundantly present.
The broad variation observed in the ratios of rotavirus RNA and infectious particles demonstrates the importance of detecting infectious viruses instead of viral RNA for the purposes involving estimations of public health risks.
通过饮用水消费评估轮状病毒的公共卫生风险,开发并优化了细胞培养-聚合酶链反应(PCR)检测方法,用于检测饮用水生产中自然污染水源中传染性环境轮状病毒株。
通过优化的细胞培养-PCR 检测方法,以不同样本量中复制病毒的存在与否为依据,通过最可能数估计传染性轮状病毒浓度。在 12 个水源样本中,有 11 个样本中检测到浓度从 0.19(0.01-0.87)到 8.3(1.8-34.0)的传染性轮状病毒,每升可检测到的感染性 PCR 单位(IPDU/l),与这些样本中传染性肠病毒的浓度无显著差异。
在 55%的样本中,轮状病毒基因组的丰度比传染性轮状病毒颗粒高 1000 至 10000 倍(3 对数-4 对数),而在其余 45%的样本中,轮状病毒基因组的丰度低于 1000 倍(<3 对数)。
轮状病毒 RNA 和传染性颗粒比例的广泛变化表明,为了评估公共卫生风险,检测传染性病毒而不是病毒 RNA 非常重要。