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LYNOAMP BC gene amplification reagent (Sysmex) was added to the supernatant, and CK19 mRNA in each lysate was amplified and detected with the RD-100i system (Sysmex), an automated molecular detection system that uses the RT-LAMP method.
In this paper, we describe the development of a molecular detection system, based on DNA array technology, for rapid and efficient detection of these vascular wilt pathogens.
Blepharisma japonicum is easily identified by a bright purple photopigment and is therefore a useful organism for developing and evaluating a molecular detection system.
Two microlitres of supernatant was analysed using the RD-100 i System (Sysmex Corporation), an automated molecular detection system that uses a reverse transcription loop-mediated isothermal amplification method (Notomi et al, 2000), and the LynoampBC Kit (Sysmex Corporation).
A total of 2- μl supernatant was analysed with the RD-100i System (Sysmex), an automated molecular detection system that uses a reverse transcription loop mediated isothermal amplification method (Notomi et al, 2000), and the LynoampBC Kit (Sysmex).
A 2-μL supernatant was analyzed with the RD-100i System (Sysmex), an automated molecular detection system that uses a reverse transcription loop-mediated isothermal amplification method, 18 and the LynoampBC Kit (Sysmex).
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Thus, the molecular detection systems had a higher level of sensitivity than standard screening of intermediate hosts by cercarial shedding when DNA was purified from pools of snails collected from endemic areas.
Despite the demonstrated advantages, the limited data on the performance, role and value of rapid culture, DST and molecular detection systems, together with concerns of increased cost and contamination rates relative to conventional culture on solid media, have dampened interest and progress in implementing these systems in low to middle income settings.
Molecular characterization of new full-length sequences will be needed to investigate the real genetic diversity of KIs-V and to help design optimized molecular detection systems.
A better knowledge of the genetic diversity of these newly discovered viruses will enable development of improved molecular detection systems and their subsequent use in epidemiologic studies involving diverse human cohorts.
The described application serves as a model for the development of rapid, molecular detection systems that offer an unprecedented combination of specificity, high throughput capabilities and robust target quantification.
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