Article

PCR up close

Published on February 7, 2021 | 10 min read
Closeup of DNA helix

Elevating PCR results

In today’s cost-conscious healthcare environment, consistent and reliable results are crucial for laboratory success.

Despite being relatively straightforward, PCR testing is not impervious to error—missteps can result in poor sensitivity and/or specificity. Every step in the PCR process must be rigorously controlled and assessed, properly calculating and applying precise denaturation and annealing temperatures during cycling to determine appropriate primer and probe concentrations for your desired targets, and more.1

Given the high degree of precision required to maintain robust laboratory processes, optimization is crucial to delivering reliable results. Labs that invest time and resources in a proper PCR setup are more likely to achieve reliable, reproducible, and robust results with higher specificity and sensitivity.

Closeup of a hand in white glove holding a sample

Maintaining result integrity

Accurate and reliable results lead to better outcomes all round, from medical research to patient diagnosis and care. Ensuring high-quality output requires precision throughout. Sample collection, transportation, sorting and preparation, storage, and QA procedures all need to be considered.

The quality and purity of samples is a key factor. Low-quality samples and contamination often compromise the end result, forcing labs to spend valuable resources correcting mistakes—this can include retraining laboratory and clinical staff (doctors, nurses etc.) Failure can be time-consuming and cost intensive. But with rigorous quality control and troubleshooting, you can produce highly accurate results every time.

To maintain result integrity and produce meaningful and reliable data at scale, there are certain performance checks which must take place:

  • Perform quality control during collection, transportation, and preparation2
  • Use correct labelling and readable barcodes when sorting samples
  • Ensure DNA/RNA are of the highest quality, including well-formulated reagents and properly designed primers and probes
  • Prevent contamination by minimizing manual tasks with end-to end automation

Today’s technology makes it possible to address these challenges and significantly improve the quality of PCR results. Combining Roche’s fully automated, high throughput analyzers with pre- and post-analytical solutions can create a fully automated workflow that can help protect against costly errors, and operate with efficiency and speed.

Key components for optimizing PCR

When it comes to PCR optimization, primers, probes, and the level of automation are of paramount importance.3

Selecting the right primers and probes

Primer and probe designs have a significant influence on the success of PCR analyses,3 and are critical for reliable and accurate quantification.

Primers and probes must be of the right quality and purity to meet the criteria necessary for optimization. The detection of specific targets requires well-designed primers and probes to ensure related non-target sequences do not compromise results.

Roche has continually innovated and developed our assay design for high sensitivity and specificity, to get the best PCR results.

The role of automation

As pressure to maximize efficiency continues to grow, advanced automation is helping to address the needs of a resource-constrained healthcare system.

Human error and bias can have a negative impact. However, automation can eliminate repetitive manual tasks, allowing labs to minimize errors and consistently deliver reliable results, faster and more efficiently, while freeing up technicians for higher-value tasks.

Roche’s industry-leading systems are designed to automate, consolidate, integrate, and standardize the entire PCR workflow, empowering labs to unlock efficiency, elevate performance, and embrace their full potential.

Roche’s commitment to advancing PCR

From the initial investment in PCR in the early 1990’s until today, Roche has been at the forefront of PCR progress. Explore Roche’s industry-defining innovations and see our vision for the future of this powerful technology.

Female lab technicians loading test tubes

What is PCR?

Polymerase chain reaction (PCR), one of the most important scientific advances of the 20th century, is an efficient and cost-effective way to copy or “amplify” small segments of DNA or RNA.
3D DNA helix

Innovation in PCR

PCR has become an indispensable tool for many applications in scientific research and clinical investigations due to the continuous development of the ecosystem surrounding the technology.

Contact us

Do you have questions about our products or services? We’re here to help. Contact a Roche representative in your region.

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Contributors

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Roche Diagnostics

Roche Diagnostics is a division of Roche, developing and integrating diagnostic solutions that address today’s healthcare challenges while anticipating tomorrow’s needs. In more than 100 countries, we provide one of the industry’s most comprehensive in vitro diagnostics portfolios spanning molecular diagnostics, clinical chemistry and immunoassays, tissue diagnostics, Point of Care testing, patient self-testing, next-generation sequencing, laboratory automation and IT, as well as digital health and decision-support solutions.

Our articles are authored by Roche Diagnostics subject matter experts, drawing on collective expertise across multiple disciplines to provide reliable insights for healthcare professionals worldwide.

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References

  1. Primer Digital. General Guidelines for PCR Optimization. https://primerdigital.com/pcr.html. Accessed 24 September 2024.
  2. Labome. Assay Development: 5 Considerations and 8 Fundamentals. https://www.labome.com/method/How-to-Develop-Assays-5-Considerations-and-8-Fundamentals.html. Accessed 24 September 2024.
  3. Wadle S, Lehnert M, Rubenwolf S, Zengerle R, von Stetten F. Real-time PCR probe optimization using design of experiments approach. Biomol Detect Quantif. 2015;7:1-8.