Evaluation of radioactive and nonradioactive gene probes and cell culture for detection of poliovirus in water samples

Abstract

Five nonradioactive probe assays were evaluated by using chemiluminescent and colormetric signals, along with two isotopic assays and cell culture, for the detection of poliovirus in concentrated water samples. In environmental samples, a 100% correlation existed between digoxigenin and single-stranded [32PIRNA probes. All probe assays detected more positive samples than the cell culture did.

Chapter 9 The Detection of Waterborne Viruses

“…Gene probes were the first approach made in the molecular biological detection of enteric viruses, and have been widely used (Dubrou et al, 1991;Enriquez et al, 1993;Margolin et al, 1993;Moore and Margolin, 1993). However they lack sensitivity and they have largely been superseded.…”

Nucleic Acid Hybridization

“…While it is feasible on paper to evaluate your research needs against these criteria, the decision must usually be determined at the lab bench. In lieu of the need to test individual systems, several studies compared the sensitivity of nonradioactive probes to the 32 P gold standard: Yang et al (1999), Plath, Peters, and Einspanier (1996), Nass and Dickson (1995), Moore and Margolin (1993), Puchhammer-Stoeckl, Heinz, and Kunz (1993), Engler-Blum et al (1993, Bright et al (1992), Kanematsu et al (1991), Lion and Haas (1990), Jiang, Estes, and Metcalf (1987), Tenberge et al (1998), Holtke et al (1992), Pollard-Knight et al (1990), Hill and Crampton (1994), Dubitsky, Brown, andBrandwein (1992), andNakagami et al (1991).…”

Detection of Fastidious Infectious Enteric Viruses in Water

“…The use of gene probes for virus diagnostic in clinical samples has been widely described (1-3). In the case of environmental water samples, the use of molecular hybridization techniques has also been reported (4,5), although the sensitivity of the method is not high enough to detect the low number of viral particles sometimes present in such samples. An alternate and more sensitive method for viral detection is the amplification of viral nucleic acids by the polymerase chain reaction (6) whose application to environmental samples is increasing ever since (7,8).…”