Optimising High-throughput Pathogen Detection
healthcaretechoutlook

Optimising High-throughput Pathogen Detection

Healthcare Tech Outlook | Friday, August 26, 2022

Centralised testing delivers greater efficiency when testing very high volumes of samples. Many diagnostic labs substantially improved their capability for reverse transcription quantitative PCR (RT-qPCR) during the pandemic. Selecting a master mix that guarantees high performance and clarity in automatic workflows is pivotal.

FREMONT, CA: While both centralised and decentralised molecular diagnostics (MDx) testing models play an essential role in patient care, centralised testing delivers greater efficiency when testing very high volumes of samples. Many diagnostic labs substantially improved their capability for reverse transcription quantitative PCR (RT-qPCR) during the pandemic to service massive PCR testing for the virus. As this testing trend winds down, these laboratories will have a prospect to repurpose their RT-qPCR capacity, by executing more syndromic testing for pathogens.

For those companies looking to design new assays or enhance existing ones, selecting an RT-qPCR master mix that guarantees high performance, clarity, and stability in automatic workflows is pivotal. Below are some methods to optimise RT-qPCR workflows.

Heat-activated Enzymes

Utilizing a heat-activated reverse transcriptase and DNA polymerase allows room-temperature set-up and contains non-specific, background amplification. Reagents and tests can be queued for loading on a platform deck for more extended periods of time without jeopardizing the accuracy of the results. 

Thermostable Enzymes

Some RNA templates can be problematic to work with, such as double-stranded RNA viruses or those viruses with a secondary structure that requires heightened initial temperatures to denature. High reverse transcriptase thermostability allows cDNA synthesis to transpire at higher temperatures, leading to more significant target specificity and the ability to identify complex viruses. Thermostability facilitates template denaturation to materialize in the presence of the enzyme, bypassing a separate RNA destructuring measure.

5x Concentrated Master Mix

Utilizing a concentrated master mix enables higher test sensitivity by permitting a more significant proportion of the PCR volume to be a clinical sample. The 5x formulation also decreases the amount of packaging and demands minor energy to ship and store the product, making it environmentally friendly and affordable.

Detect RNA and DNA Templates‚Äč

Determining a single master mix including a reverse transcriptase indicates that the same master mix can be employed for the detection of bacterial and DNA and RNA viral targets. This streamlines ordering and assay set-up. Ensuring the master mix also includes an abundance of essential PCR reaction components allows the detection of multiple pathogens in one reaction. 

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