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EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay
Published on: September 11, 2016
Detection and quantification of enteroviruses in coastal seawaters from Bohai Bay, Tianjin, China
Minglu Zhang1, Huabing Zhao, Jian Yang
1Key Laboratory of Bioactive Materials, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071, China. zhangml1982@yahoo.com.cn
Abstract:
An 8-month survey was conducted to detect and quantify enteroviruses in Tianjin coastal seawaters of Bohai Bay to assess coastal water quality. Ten water samples were collected from Bohai Bay for the detection and quantification of enteroviruses by conventional reverse transcription polymerase chain reaction (RT-PCR) and SYBR Green real-time quantitative RT-PCR (qRT-PCR). Total viral nucleic acid was extracted from 500 mL of seawater samples concentrated by Centricon plus-70 centrifugal filter devices. The viral recovery rate was 29.1% based on viral seeding study. The centrifugal ultrafiltration method applied is effective for viral recovery from small volume of polluted water, which may have broader applications to monitoring human virus in aquatic environment. Our results indicated that there was a severe viral contamination in seawater of Bohai Bay. Enteroviruses were detected at concentrations ranging from 1.7 x 10(6) to 6.3 x 10(7) copies/L by qRT-PCR. Sequencing analyses identified that all of the twenty clones as poliovirus type 2. This is the first quantitative report of human viruses in coastal waters of a metropolitan city in China. This study emphasized the importance for the local and central governments to monitor and assess the water quality.
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