Isolation and detection of enterovirus RNA from large-volume water samples by using the NucliSens miniMAG system and real-time nucleic acid sequence-based amplification.

Appl Environ Microbiol

National Institute for Public Health and the Environment (RIVM), Microbiological Laboratory for Health Protection (MGB), Health-Related Water Microbiology, P.O. Box 1/Antonie van Leeuwenhoeklaan 9, 3720 BA Bilthoven, The Netherlands.

Published: July 2005

Concentration of water samples is a prerequisite for the detection of the low virus levels that are present in water and may present a public health hazard. The aim of this study was to develop a rapid, standardized molecular method for the detection of enteroviruses in large-volume surface water samples, using a concentration method suitable for the detection of infectious viruses as well as virus RNA. Concentration of water was achieved by a conventional filter adsorption-elution method and ultrafiltration, resulting in a 10,000-fold concentration of the sample. Isolation of virus RNA by a silica-based RNA extraction method was compared with the nonmagnetic and magnetic NucliSens RNA isolation methods. By using the silica-based RNA extraction method in two out of five samples, enterovirus RNA was detected, whereas four out of five samples were positive following RNA isolation with magnetic silica beads. Moreover, estimated RNA levels increased at least 100 to 500 times. Furthermore, we compared enterovirus detection by an in-house reverse transcription (RT)-PCR with a novel commercially available real-time nucleic acid sequence-based amplification (NASBA) assay. We found that the rapid real-time NASBA assay was slightly less sensitive than our in-house RT-PCR. The advantages, however, of a commercial real-time NASBA assay, like the presence of an internal control RNA, standardization, and enormous decrease in turnaround time, makes it an attractive alternative to RT-PCR.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1169005PMC
http://dx.doi.org/10.1128/AEM.71.7.3734-3740.2005DOI Listing

Publication Analysis

Top Keywords

water samples
12
nasba assay
12
rna
10
enterovirus rna
8
real-time nucleic
8
nucleic acid
8
acid sequence-based
8
sequence-based amplification
8
concentration water
8
virus rna
8

Similar Publications

Purpose: A comprehensive survey was conducted to assess the prevalence of Schistosoma japonicum infection in humans, water buffaloes, and snails in the two endemic municipalities of Talibon and Trinidad in Bohol, Philippines, which are nearing elimination.

Methods And Results: Human stool and blood samples were collected from barangays with snail breeding sites, and results showed higher positivity rates using the rSjTPx-1-ELISA compared to the Kato-Katz technique. Human stool examination for showed a 0.

View Article and Find Full Text PDF

Background: This study aimed to evaluate the efficacy of polydopamine (PDA) functionalization on orthodontic brackets in inhibiting biofilm formation and promoting surface bioactivity to buffer the acidity of caries-causing bacteria around orthodontic brackets and prevent demineralization. The stability of the coating in artificial saliva (AS) and distilled water was evaluated, along with its effect on pH changes in simulated body fluid (SBF) and distilled water.

Methods: Maxillary incisor orthodontic brackets underwent PDA functionalization using a dopamine hydrochloride solution following a specific protocol.

View Article and Find Full Text PDF

Legacy and emerging Organophosphate flame retardants (OPFRs) in water and sediment from the Pearl River Delta to the adjacent coastal waters of the South China Sea: Spatioseasonal variations, flux estimation and ecological risk.

Environ Pollut

January 2025

Guangdong-Hong Kong-Macao Joint Laboratory for Contaminants Exposure and Health, School of Environmental Science and Engineering, Institute of Environmental Health and Pollution Control, Guangdong University of Technology, Guangzhou, 510006, China. Electronic address:

The industrialization and urbanization along the Pearl River Delta (PRD) have exacerbated the issue of pollution in aquatic environments by organophosphate flame retardants (OPFRs). Historical cumulative pollution from legacy OPFRs, combined with newly emerging OPFRs, has increased the severity and complexity of OPFR pollution in this region. We explored the contamination profile, input flux and risk of legacy and emerging OPFRs in surface waters and in sediment samples of the PRD.

View Article and Find Full Text PDF

Despite their potential risks to human health and the environment at ng/L to μg/L concentrations, there has been relatively little effort to measure trace organic compounds (TOrCs) in surface waters of Central America. The concentrations of eighteen TOrCs detected at eleven surface water sites in the Lempa River basin of El Salvador and four sources of drinking water for the cities of San Salvador, Antiguo Cuscatlán, Soyapango, and Santa Tecla are reported here. All samples were analyzed via liquid chromatography with tandem mass spectrometry (LC-MS/MS).

View Article and Find Full Text PDF

The potential of urinary miR-200c-3p as a biomarker of fluorosis in rats.

Ecotoxicol Environ Saf

January 2025

Key Lab of Etiology and Epidemiology, Education Bureau of Heilongjiang Province & Ministry of Health (23618504), Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, Harbin, Heilongjiang Province 150081, China. Electronic address:

Fluorine is a strong oxidizing element and excessive intake can have harmful effects, particularly on the body's calcified tissues. Recent studies have demonstrated a link between miRNA and fluorosis. This study aimed to evaluate the time-dose-effect relationship of miR-200c-3p in plasma, urine and cartilage of rats with drinking water fluorosis, and to explore its potential as a biomarker.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!