Development of an effective method for recovery of viral genomic RNA from environmental silty sediments for quantitative molecular detection

Takayuki Miura, Yoshifumi Masago, Daisuke Sano, Tatsuo Omura

研究成果: Article

19 引用 (Scopus)

抜粋

Nine approaches to recover viral RNA from environmental silty sediments were newly developed and compared to quantify RNA viruses in sediments using molecular methods. Four of the nine approaches employed direct procedures for extracting RNA from sediments (direct methods), and the remaining five approaches used indirect methods wherein viral particles were recovered before RNA extraction. A direct method using an SDS buffer with EDTA to lyse viral capsids in sediments, phenol-chloroform-isoamyl alcohol to extract RNA, isopropanol to concentrate RNA, and magnetic beads to purify RNA resulted in the highest rate of recovery (geometric mean of 11%, with a geometric standard deviation of 0.02; n = 7) of poliovirus 1 (PV1) inoculated in an environmental sediment sample. The direct method exhibiting the highest rate of PV1 recovery was applied to environmental sediment samples. One hundred eight sediment samples were collected from the Takagi River, Miyagi, Japan, and its estuary from November 2007 to April 2009, and the genomic RNAs of enterovirus and human norovirus in these samples were quantified by reverse transcription (RT)-quantitative PCR (qPCR). The human norovirus genome was detected in one sample collected at the bay, although its concentration was below the quantification limit. Meanwhile, the enterovirus genome was detected in two samples at the river mouth and river at concentrations of 8.6 × 102 and 2.4 × 102 copies/g (wet weight), respectively. This is the first report to obtain quantitative data for a human pathogenic virus in a river and in estuarine sediments using RT-qPCR.

元の言語English
ページ(範囲)3975-3981
ページ数7
ジャーナルApplied and environmental microbiology
77
発行部数12
DOI
出版物ステータスPublished - 2011 6

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

フィンガープリント Development of an effective method for recovery of viral genomic RNA from environmental silty sediments for quantitative molecular detection' の研究トピックを掘り下げます。これらはともに一意のフィンガープリントを構成します。

  • これを引用