Sentence examples for simplex format from inspiring English sources
Based on the facts that φA1122 [7], [49], [51] and L-413C [52], [53], [53] can lyse almost all (99.8 99.9%) Y. pestis strains of several thousand tested, we propose a broad-range qPCR method in a simplex format.
In our experience, the simplex format strategy (i.e. based on testing phages and viral pathogens in distinct amplification reactions) proved to be the most simple and cost-effective for routine molecular diagnosis since it does not require the specific development of multiplex reactions and relies on a unique control reaction for DNA viruses and another for RNA viruses.
Reactions were carried out in simplex format in 25 μl containing 200 ng DNA, 1× TaqMan® Universal PCR Master mix (Applied Biosystems), 900 nM each primer and 200 nM probe.
Conversely, only 6 isolates could be processed per run if the same test was to be carried out in conventional simplex pyrosequencing format (i.e., one well per pyrosequencing reaction).
It was validated for both PCR and RT-PCR real time techniques, in simplex or multiplex format and, in the case of RT-PCR assays, one-step or two-step amplification formats.
The criteria to be addresses regarding ICs were: (i) to monitor all steps of the diagnostic procedure (extraction, RT, PCR); (ii) to be amenable for DNA and RNA viruses ; (iii) to rely on a detection system specific of the phage(s) to avoid complex molecular constructs; (iv) to be usable in either simplex or multiplex format, and in one-step or two-step RT-PCR reactions.
All primer pairs produced virus-specific fragments in both simplex and multiplex formats, thus confirming the efficacy of the newly developed mRT-PCR for detection of these viruses.
When compared to the two simplex reactions, the duplex format did not shift Ct values and did in particular not compromise the sensitivity of the two assays.
The multiplex format is advantageous over simplex assays as it is particularly economical in terms of cost of material (mastermix, consumables) and turnaround time and suitable for small volume samples and high-throughput.
To date, a combinatory matrix-based approach based on simplex (eventually duplex or multiplex) qPCR methods in a 96-well (or larger) format could also be envisaged for such purposes as long as within the global GMP Universe, common elements (such as the p35S, the GM-traits…) are being applied in the transformation vectors.
