Study On The Concentration Method And PCR Detection Of Enteroviruses From Environmental Waters

At present, infection of pathogens spreaded by water is the most serious problem in the world, which has caused many epidemics, especially enteroviruses which caused epidemics more than any other viruses, and also the impacted area is the largest. So, study on the enteroviruses become a research hotspot. However, the concentration of the enteroviruses in waters are always very low, and can not be detected directly. Enteroviruses must be concentrated from water to achieve the detection level. Besides, traditional detection method of cell cuture is very complicated, and can not be used universally. Therefore, it is very useful for study on enteroviruses from waters to choose a highly effective concentration method and a sensitive detection method.In this paper, we summarized all the concentration methods used, and we carefully chose four methods to compare,which were microporous filter adsorption-elution method, talcum powder-diatomaceous earth adsorption layer absorption-elution method and talcum powder-diatomaceous earth flocculation precipitation method. Considering about the virus recovery and reliability of the methods, we take the microporous filter adsorption-elution method for the best one. Additionally, we also summarized all the detection methods, and emphasized on the introduction of the principles of PCR and its application. Based on RT-PCR, we build the nested PCR method to detect enteroviruses in waters by universal primer, which amplificated the RT -PCR amplification, and this method could greatly increase the specificity and sensitivity. Furthermore, we deteced the typical surface water in Xi'an city consecutively by nested PCR, and made a analysis of the distrbution characteristic of enteroviruses in Xi'an city.