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Description
| - Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method and has the potential to compete with PCR because of its simplicity, rapidity, specificity, and cost-effectiveness. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay has emerged as a powerful gene amplification tool for the rapid identification of microbial infections and is being increasingly used for rapid detection and typing of emerging viruses, for example the Chikungunya virus, West Nile virus, dengue virus etc. The detection of gene amplification can be accomplished by either agarose gel electrophoresis or by real-time monitoring in the turbidimeter. Moreover, the gene amplifification can be eye-visualized either by formation of the white precipitate or by using fluorescent intercalating dye (e.g. SYTO) and UV lamp. Because the LAMP method uses a Bst DNA polymerase long fragment, the reaction can be carried out under isothermal conditions in the water bath or thermal block. For these reasons this method is very suitable for detection in the field conditions as it doesn´t require any sophisticated equipment. We introduced one-step, single-tube RT-LAMP assay as well as RT-qPCR assay for the detection of the Chikungunya virus. The sensitivity of the RT-LAMP assay targeting E1gene of the Chikungunya virus genome was compared with the CHIKV-specific RT-qPCR assay. Our results indicate that RT-LAMP is more sensitive method in contrast to RT-qPCR. On the other hand RT-qPCR technique is more robust than RT-LAMP.
- Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method and has the potential to compete with PCR because of its simplicity, rapidity, specificity, and cost-effectiveness. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay has emerged as a powerful gene amplification tool for the rapid identification of microbial infections and is being increasingly used for rapid detection and typing of emerging viruses, for example the Chikungunya virus, West Nile virus, dengue virus etc. The detection of gene amplification can be accomplished by either agarose gel electrophoresis or by real-time monitoring in the turbidimeter. Moreover, the gene amplifification can be eye-visualized either by formation of the white precipitate or by using fluorescent intercalating dye (e.g. SYTO) and UV lamp. Because the LAMP method uses a Bst DNA polymerase long fragment, the reaction can be carried out under isothermal conditions in the water bath or thermal block. For these reasons this method is very suitable for detection in the field conditions as it doesn´t require any sophisticated equipment. We introduced one-step, single-tube RT-LAMP assay as well as RT-qPCR assay for the detection of the Chikungunya virus. The sensitivity of the RT-LAMP assay targeting E1gene of the Chikungunya virus genome was compared with the CHIKV-specific RT-qPCR assay. Our results indicate that RT-LAMP is more sensitive method in contrast to RT-qPCR. On the other hand RT-qPCR technique is more robust than RT-LAMP. (en)
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Title
| - The introduction of RT-LAMP method and comparison of its sensitivity with RT-qPCR in the detection of Chikungunya virus
- The introduction of RT-LAMP method and comparison of its sensitivity with RT-qPCR in the detection of Chikungunya virus (en)
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skos:prefLabel
| - The introduction of RT-LAMP method and comparison of its sensitivity with RT-qPCR in the detection of Chikungunya virus
- The introduction of RT-LAMP method and comparison of its sensitivity with RT-qPCR in the detection of Chikungunya virus (en)
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skos:notation
| - RIV/70565813:_____/13:#0000291!RIV14-MV0-70565813
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http://linked.open...avai/riv/aktivita
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http://linked.open...avai/riv/aktivity
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http://linked.open...vai/riv/dodaniDat
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http://linked.open...aciTvurceVysledku
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http://linked.open.../riv/druhVysledku
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http://linked.open...iv/duvernostUdaju
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http://linked.open...titaPredkladatele
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http://linked.open...dnocenehoVysledku
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http://linked.open...ai/riv/idVysledku
| - RIV/70565813:_____/13:#0000291
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http://linked.open...riv/jazykVysledku
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http://linked.open.../riv/klicovaSlova
| - pathogen detection; qPCR; Loop-mediated isothermal amplification (en)
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http://linked.open.../riv/klicoveSlovo
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http://linked.open...ontrolniKodProRIV
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http://linked.open...v/mistoKonaniAkce
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http://linked.open...i/riv/mistoVydani
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http://linked.open...i/riv/nazevZdroje
| - 26. KONGRES ČESKOSLOVENSKÉ SPOLEČNOSTI MIKROBIOLOGICKÉ
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http://linked.open...in/vavai/riv/obor
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http://linked.open...ichTvurcuVysledku
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http://linked.open...cetTvurcuVysledku
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http://linked.open...vavai/riv/projekt
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http://linked.open...UplatneniVysledku
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http://linked.open...iv/tvurceVysledku
| - Procházková, Jiřina
- Bílek, Karel
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http://linked.open...vavai/riv/typAkce
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http://linked.open.../riv/zahajeniAkce
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number of pages
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http://purl.org/ne...btex#hasPublisher
| - Československá společnost mikrobiologická
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https://schema.org/isbn
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