Zhang Ming-Lu, Yang Jian, Zhao Hong, Zhu Lin, Zhao Hua-Bing, Cai Bao-Li
Key Laboratory of Bioactive Materials, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071, China.
Huan Jing Ke Xue. 2009 Jun 15;30(6):1608-13.
Hepatitis A virus (HAV) is single-strainded RNA virus that causes infectious hepatitis A. Detection and quantification of hepatitis A virus in Tianjin coastal seawater of Bohai Bay were carried out by conventional RT-PCR and SYBR Green real-time quantitative RT-PCR using the primers based on the conserved sequence at the VP1-VP2 genes of HAV. The nine samples were taken at Tianjin coastal seawater of Bohai Bay locating in the south of Tanggu, in summer, autumn and winter of 2007 and spring of 2008. For viral detection, seawater samples were concentrated either using a small ultrafiltration system (Millipore Pellicon Mini TFF) or a Centriprep-100 centrifugal ultrafiltration device (Millipore Centricon Plus-70). RT-PCR analysis showed that a 192 bp HAV cDNA was amplified from all nine seawater samples and the sequence identities of these cDNAs to the homologous sequence in the GenBank were between 95% and 100%. SYBR Green real-time quantitative RT-PCR analysis indicated that HAV concentration in these samples ranged from 5.35 x 10(6) to 4.51 x 10(7) virus particles/L.
甲型肝炎病毒（HAV）是一种引起甲型传染性肝炎的单链RNA病毒。采用基于甲型肝炎病毒VP1-VP2基因保守序列设计的引物，通过常规逆转录聚合酶链反应（RT-PCR）和SYBR Green实时定量逆转录聚合酶链反应，对渤海湾天津沿海海水中的甲型肝炎病毒进行检测和定量分析。于2007年夏、秋、冬及2008年春在塘沽以南的渤海湾天津沿海海水采集了9份样品。为进行病毒检测，海水样品采用小型超滤系统（密理博Pellicon Mini TFF）或Centriprep-100离心超滤装置（密理博Centricon Plus-70）进行浓缩。RT-PCR分析显示，从所有9份海水样品中均扩增出一条192 bp的甲型肝炎病毒cDNA，这些cDNA与GenBank中同源序列的序列同一性在95%至100%之间。SYBR Green实时定量RT-PCR分析表明，这些样品中的甲型肝炎病毒浓度范围为5.35×10⁶至4.51×10⁷病毒颗粒/升。
