• Korean Journal of Microbiology
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• v.35 no.1
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• pp.41-46
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• 1999

In detecting pathogeuic viruses in water sample, polymerase-chain-reaction (PCR) amplification was uscd. In order lo obtain the intact viral particlc, five concentration techniques were compared and an improved procedure was developed with some modifications. Among them, adsorption-elution~EG precipitation and flocculatio~~iultracentriEugation were more efficient than others with thc detection limit of 10 PFU $ml^{-1}$. By the additional step removing inhibitory compounds for PCR reaction, the purity of the concentrated sample was improved and the detection limit was lowered by one order (to 1 PFU $ml^{-1}$. To examine the availability of the optimized procedure for field surveys, the distributions of enterovirus in Han River were estimated using the novel procedure. Seventy-five percentage (618) of sewagc samples and twenty percentage (2110) of river water samples were positive for enterovirus. These results indicate that adsorption-elutionPEG precipitation by PCR method is useful for the prompt and handy monitoring of viral contaminaiton in water environment and pathogenic viruses are widely distributed in water environments of Seoul.

• Food Science and Biotechnology
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• v.18 no.5
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• pp.1150-1154
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• 2009

Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.