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Enabling shorter turnaround times in molecular PoC with the right reagents

Reagents designed to perform wherever and whenever answers are needed

Rapid onsite analyses to support diagnosis and treatment decisions have become a cornerstone of medicine in our globalized society. Molecular Point-of-care testing (POCT) not only extends the reach of modern medicine to broader populations, but also underpins our understanding and management of public health.

Assays for molecular PoC must work reliably, delivering accurate results under variable handling and environmental conditions. They must also be easy to transport, store and use. Finally, they must be manufactured quickly, safely and cost-effectively. Using reagents designed for the specific conditions of PoC is key to achieving these requirements.

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The value of PoC testing lies in delivering on-the-spot results for treatment decisions.

Turnaround can be shortened by boosting assay speed, especially in the case of RT-PCR and PCR-based tests.

KAPA3G, designed for speed and robustness, can perform a full PCR within only 23 minutes (all polymerases were used according to manufacturer's instructions, data on file at Roche).

The protocols may become even longer in the case of RT-PCR because of the additional reverse transcription step. Fortunately, NxtScript 2G does it very fast and completes reverse transcription in only 10 minutes. And what’s more, NxtScript 2G is a heat-powered reverse transcriptase evolved toward high thermostablity. It performs reverse transcription at high temperatures (up to 70°C) to easily tackle any difficult RNA target.

 

 

 

Fast cycling shortens assay time

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Enhanced multiplexing capacity

Another strategy to shorten turnaround time is to enable the simultaneous detection of multiple analytes in one reaction: multiplexing. KAPA3G HotStart DNA polymerase was evolved for exceptional speed and tolerance. It has also demonstrated a highly robust multiplexing capacity.

 

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Robust to variation and inhibitors


Sample preparation is often time-consuming and variable. Thus, developers of PoC diagnostics seek to simplify – if not completely eliminate – sample processing. As a result, analyses may be exposed to PCR-inhibiting substances naturally contained in biological matrices. Roche CustomBiotech enzymes, such as KAPA3G, have been tested against a broad range of inhibitors inherent to liquid biopsies, tissues, and standard sample preparation methods, and have demonstrated robust performance even with low-processed samples.

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Roche CustomBiotech enzymes have been tested with a broad range of inhibitors inherent to liquid biopsies, tissues or standard sample preparation methods.
 

Tolerance is defined as a shift in Cp of ≤ 3 and  a reduction in fluorescence of ≤ 50% of the control total fluorescence.

One-stop shop: Roche CustomBiotech lyo-ready reagents

With a view towards demands of manufacturers of modern diagnostics, Roche CustomBiotech continues to expand and improve its portfolio of high-quality enzymes and master mixes. Our two major families of reverse transcriptases – NxtScript for RNA and HawkZ05 Fast for both RNA and DNA – include lyo-ready formulations.
For our DNA polymerases, lyo-ready options include aptamer-based, and antibody-based hot start. Many of these DNA polymerases and reverse transcriptases can be combined without buffer modifications to create fast, multiplexed, inhibitor-tolerant, dried-down one-step RT-PCR assays. Plus, our commitment to setting the pace for PoC diagnostics does not stop at enzymes and master mixes.
Roche CustomBiotech delivers a broad spectrum of premium ingredients at scale and on time for assays used today and those to come tomorrow.

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Regulatory disclaimers are listed on the respective product pages.