Reagents Ideal for High-Throughput Multiplex Assays
High-throughput platforms, when combined with multiplexing capabilities, offer a formidable toolkit for diagnosticians, enhancing the depth of information obtained from each sample. Molecular multiplex assays can significantly lower the cost of testing multiple targets at one time. Compared to singleplex reactions, multiplex requires less reagents, less time, and less labor to achieve the same result. However, in mutiplex assays, the inherent variability between the physicochemical characteristics (e.g. length, GC content) of the amplified sequences , flanking regions and secondary structures can impact Ct values and create an imbalance in the amplification of certain targets. Meridian’s enzymes and master mixes are formulated for robust amplification under a wide range of temperatures, reaction and cycling conditions, and specimen types (blood, saliva, urine, and stool). They are ideal for multiplexing assays, which typically require diverse reaction conditions, and ensure that amplification is consistent across all the assay targets, allowing for accurate and reliable results for each analyte.
A. 55C MMLV-RT (MDX117) exhibits robust performance in multiplexed assays with samples containing sputum and Universal Transport Media (UTM)
Flu A
MERS-CoV
RSV
55C MMLV-RT
Performance of 55C MMLV-RT in multiplex RT-qPCR. 55C MMLV-RT was tested in a triplex one-step RT-qPCR assay on respiratory RNA virus targets (Influenza A, MERS-CoV and RSV). The results demonstrate the sensitivity and reproducibility of 55C MMLV-RT even in challenging conditions such as multiplex RT-qPCR reactions where targets may differ greatly in abundance.
B. Lyo-Ready ™ Direct DNA qPCR Saliva (MDX132) demonstrates sensitive detection in multiplex assays with samples containing sputum
Mycoplasma pneumoniae
ADV
CMV
0% Artificial sputum | 10% Artificial sputum | 20% Artificial sputum | 60% Artificial sputum
Performance of Lyo-Ready ™ Direct DNA qPCR Saliva. Mycoplasma pneumoniae genomic DNA, adenovirus (ADV) and cytomegalovirus (CMV) were amplified in a triplex qPCR assay in the presence of 0% ( Black ), 10% ( red ), 20% ( blue ) and 60% ( green ) artificial sputum. The results illustrate that the performance of the Lyo-Ready ™ Direct DNA qPCR Saliva is not affected by increasing concentrations of sputum.
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