关键词: 氯化钠-氯化铝沉淀法；滤膜吸附法；肠道病毒；逆转录PCR；巢式PCR

Abstract:

Effective enrichment and recovery of virus particles from water is the primary task in virus detection in water samples. The present study used sodium chloride-aluminum chloride precipitation method and cation-coated filter method to concentrate polioviruses from a simulated virus contaminated water sample and a real untreated water sample collected from a sewage treatment plant in Shanghai. RNA was extracted from concentrated viruses and then reverse transcription-PCR (RT-PCR) and nest PCR were perfomed. The results revealed that polioviruses could be detected from the real water sample by means of two concentration methods, indicating that viruses had been effectively recovered from water by those two methods. The presence of polioviruses was detected from virtual volume of 3.5 mL untreated water sample using sodium chloridealuminium chloride precipitation method and from virtual volume of 2.1 mL untreated water sample using cation-coated filter method. We also found the presence of polioviruses in the influent and effluent water samples collected from another three sewage treatment plants using sodium chloridealuminium chloride precipitation method and RT-nest-PCR detection. In addition, a recombinent plasmid containing poliovirusspecific fragment was constructed and prepared as a positive experimental control for the further poliovirus detection. Our study provided preliminary data for applications of the two different methods for virus concentration in water samples and observed potential enterovirus pollution in aquatic environment in Shanghai.

Key words: NaCl-AlCl₃ precipitation method; cation-coated filter method; enterovirus; reverse transcription-PCR (RT-PCR);nest PCR