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The real-time assay described is rapid, specific and sensitive.
The multiplex real-time assay was at least 10 times more sensitive than conventional protocols for viroid and phytoplasma detection.
The real-time assay did not amplify sequences from the corresponding genomic regions of feline calicivirus (also in the genus Vesivirus) and representative members of the genus Norovirus.
The multiplex real-time assay developed in this study can be used as a valuable tool to detect MDR virulent Salmonella, thus enhancing the safety of food.
The real-time assay, optimised for the simultaneous detection of blaVIM and blaKPC, identified 3 and 12 isolates positive for both blaVIM/blaKPC and for blaKPC, respectively.
This TaqMan real-time assay was tested using four different reagent kits and was shown to be robust and stable, with no significant differences in sensitivity between kits.
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The sensitivity of each real time assay was established by comparing real time PCR to bacterial culture.
To confirm successful DNA extraction a human C reactive protein real time assay was run for each DNA extraction.
The real time assay sequences, cycling conditions and quality control process are described in the Supplementary Text S1 and Supplementary Figure S1.
This is a non-real time assay, which means key cellular events can be missed.
Real Time assay oligos for these genes are described in Table 4.
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