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Development of a Virus Concentration Method and its Application for the Detection of Noroviruses in Drinking Water in China  

Liu, Junyi (Wuhan Institute of Virology, Chinese Academy of Science)
Wu, Qingping (Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology)
Kou, Xiaoxia (Wuhan Institute of Virology, Chinese Academy of Science)
Publication Information
Journal of Microbiology / v.45, no.1, 2007 , pp. 48-52 More about this Journal
Abstract
A new procedure for the concentration of nonoviruses from water samples has been developed. This procedure (calcium flocculation-citrate dissolution method) uses the following steps: virus flocculation formed by treatment with 1 M $CaCl_2$ and 1 M $Na_2HPO_4$, virus release by sodium citrate dissolution (0.3 M Na citrate, pH 3.5), and virus re-concentration by ultrafiltration. When reverse transcription (RT)-PCR was performed after the procedure, the overall detection sensitivity for seeded noroviruses in a one liter drinking water sample was as low as 1 RT-PCR unit, which is equal to a $10^{-6}$ dilution of the virus sample. This approach showed at least a 5-fold-higher sensitivity than the current method with its three steps of adsorption-elution-concentration. The newly developed procedure was used to test different brands of bottled drinking water from China for putative contamination with noroviruses. A total of 144 samples were analyzed; all of the samples were negative for norovirus specific nucleic acids.
Keywords
Noroviruses; concentration; drinking water;
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