DETECTION OF ADENOVIRUS AND ENTEROVIRUS BY PCR AMPLIFICATION IN POLLUTED WATERS

被引:31
作者
GIRONES, R [1 ]
PUIG, M [1 ]
ALLARD, A [1 ]
LUCENA, F [1 ]
WADELL, G [1 ]
JOFRE, J [1 ]
机构
[1] UMEA UNIV,DEPT VIROL,S-90185 UMEA,SWEDEN
关键词
VIRUS DETECTION; ADENOVIRUS; ENTEROVIRUS; POLYMERASE CHAIN REACTIONS; PCR;
D O I
10.1016/0273-1223(95)00291-T
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
Several systems for virus recovery from environmental samples and extraction of nucleic acid were tested by adding adenovirus 2 and poliovirus 1 to different sewage samples. The most promising method involved: concentration of viruses by centrifugation, and elution of the pelleted viruses by treatment with 0.25 N glycine buffer pH 9.5. The nucleic acids were extracted by adsorption of RNA and DNA to silica particles. One aliquot was directly used for a two-step PCR in a nested fashion, with specific primers for all adenoviruses; the other aliquot was used to synthesize cDNA and a nested two-step PCR with specific primers for enteroviruses. The specificity and sensitivity of the selected primers were evaluated, the 47 human adenovirus serotypes were identified and 24 different enterovirus strains were-recognized. The sensitivity of the nucleic acid extraction, cDNA synthesis and nested PCR amplification was estimated to be between 1 and 10 viral particles. Sewage and polluted river samples were analyzed showing, as expected, a much higher number of positive samples by the method described than by tissue culture analysis, a high prevalence of hepatitis A virus in sewage and the adenoviruses as the most commonly detected virus in the environmental samples analyzed.
引用
收藏
页码:351 / 357
页数:7
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