An integrated cell culture and quantitative PCR assay for detection of infectious rotaviruses in environmental waters
Rotaviruses exist widely in water environments and pose great health risk. Outbreaks of viral gastroenteritis associated with the contamination of water supplies have been reported. And the occurrences of rotaviruses in water environments pose an urgent need for rapid and sensitive detection method to monitor rotaviruses in water environments,which is essential for the prevention of possible infection sources. To overcome the disadvantages of traditionally plaque assay and direct PCR method,an integrated cell culture and reverse transcription quantitative PCR (ICC-RT-qPCR) assay was applied to detect infectious rotaviruses based on detection of viral RNA during replication in cells. Only infectious rotaviruses can be detected because they the only ones that can infect cells and produce RNA. This method provides a higher sensitivity because the cell culturing prior to the nucleic acid amplification increases the amount of infectious viruses and therefore allows for detection of viruses before they produce observable plaque.In addition,the assay can decrease the impact of inhibitory compounds present in environmental samples to PCR assay because the nucleic acid is extracted from infected host cells rather than from water samples directly.
Dan Li Wan Yang Miao He Xiu-hua Hu Ran-Chang Shi
Environmental Simulation and Pollution Control (ESPC) State Key Joint Laboratory,Department of Environmental Science and Engineering,Tsinghua University,Beijing,100084,China
国际会议
International Conference on Environment Simulation and Pollution Control(第六届环境模拟与污染控制学术研讨会)
北京
英文
158-159
2009-11-01（万方平台首次上网日期，不代表论文的发表时间）