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|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Monpoeho, S
|e verfasserin
|4 aut
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| 245 |
1 |
0 |
|a Quantification of enterovirus RNA in sludge samples using single tube real-time RT-PCR
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| 264 |
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1 |
|c 2000
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| 336 |
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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| 338 |
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|a Band
|b nc
|2 rdacarrier
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| 500 |
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|a Date Completed 09.11.2000
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| 500 |
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a We have developed a quantitative RT-PCR method that can be used to determine the amount of enterovirus RNA in urban sludge samples. This method combines Taq-Man technology with the ABI Prism 7700 real-time sequence detection system. We optimized a one-step RT-PCR that uses a dual-labeled fluorogenic probe to quantify the 5' noncoding region of enteroviruses. For accurate quantification of the number of copies, a Mahoney type 1 poliovirus RNA standard was designed and produced using genetic engineering. This fragment, quantified using the Ribogreen method, was used in serial dilutions as an external standard. The method had a 7-log dynamic range (5 to 2 x 10(7)). PCR inhibitors were removed by extracting viral RNA (after virus concentration) using the RNeasy mini kit with added polyvinylpyrrolidone (PVP) and running the amplification reaction with a mixture containing PVP and T4 gene 32 protein. This real-time quantification of enterovirus RNA allows large numbers of samples to be screened. Its sensitivity, simplicity and reproducibility render it suitable as a screening method with which to characterize enteroviruses, the presence of infectious particles being subsequently confirmed by cell culture
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| 650 |
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|a Research Support, Non-U.S. Gov't
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| 650 |
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|a Technical Report
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| 650 |
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7 |
|a Fluorescent Dyes
|2 NLM
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| 650 |
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7 |
|a RNA, Viral
|2 NLM
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| 650 |
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7 |
|a Sewage
|2 NLM
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| 700 |
1 |
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|a Dehée, A
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Mignotte, B
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Schwartzbrod, L
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Marechal, V
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Nicolas, J C
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Billaudel, S
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Férré, V
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1988
|g 29(2000), 1 vom: 17. Juli, Seite 88-93
|w (DE-627)NLM012627046
|x 1940-9818
|7 nnns
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| 773 |
1 |
8 |
|g volume:29
|g year:2000
|g number:1
|g day:17
|g month:07
|g pages:88-93
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