Department of Civil and Environmental Engineering, Michigan State University, East Lansing, MI 48824, USA.
J Appl Microbiol. 2009 Oct;107(4):1219-29. doi: 10.1111/j.1365-2672.2009.04300.x. Epub 2009 Apr 9.
To assess human adenovirus (HAdV) diversity in environmental samples based on sequence comparisons of hexon gene fragments amplified using newly designed HAdV-specific polymerase chain reaction (PCR) assays.
Six PCR primer sets were designed based on 56 aligned hexon sequences from NCBI GenBank to amplify different hexon gene sections (241-349 bp) of the six HAdV species. The amplified hexon genes from wastewater samples were cloned, sequenced, and compared with those in publicly accessible databases (i.e. NCBI GenBank) by using the Blast program. A total of 46 analysed positive clones were affiliated to five HAdV serotypes, i.e. 1, 2, 12, 31 and 41. Similarities between the cloned and database hexon sequences ranged from 95.9 to 100% (with an average of 98.1 +/- 1.0%).
The designed primers showed higher amplification efficiencies when compared with the existing assays. Using the new assays, HAdV species A, C, and F (serotypes 1, 2, 12, 31 and 41 in particular) were identified in the studied municipal wastewater.
The six PCR primer sets developed in this study can be used to efficiently amplify hexon gene fragments in HAdV. Multiple HAdV serotypes identified in the municipal wastewater provide new information about HAdV diversity in environmental samples.
通过比较使用新设计的人腺病毒（HAdV）特异性聚合酶链反应（PCR）检测方法扩增的六邻体基因片段的序列，评估环境样本中的 HAdV 多样性。
根据来自 NCBI GenBank 的 56 个对齐六邻体序列，设计了 6 个 PCR 引物组，以扩增六种 HAdV 物种的不同六邻体基因部分（241-349 bp）。从废水样本中扩增的六邻体基因被克隆、测序，并通过 Blast 程序与公共数据库（即 NCBI GenBank）中的基因进行比较。总共分析了 46 个阳性克隆，它们与五种 HAdV 血清型有关，即 1、2、12、31 和 41。克隆和数据库六邻体序列之间的相似性范围为 95.9%至 100%（平均为 98.1 +/- 1.0%）。
与现有检测方法相比，设计的引物显示出更高的扩增效率。使用新的检测方法，在研究的城市废水中鉴定出人腺病毒 A、C 和 F （特别是血清型 1、2、12、31 和 41）。
本研究中开发的 6 个 PCR 引物组可用于有效地扩增 HAdV 的六邻体基因片段。在城市废水中鉴定出的多种 HAdV 血清型为环境样本中的 HAdV 多样性提供了新的信息。
