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Related Experiment Videos

[Present status and future on automated PCR testing for microorganisms]

T Hirose1

  • 1Diagnostic Division, Nippon Roche K.K., Tokyo.

Rinsho Byori. the Japanese Journal of Clinical Pathology
|November 1, 1996
PubMed
Summary

Polymerase Chain Reaction (PCR) technology has advanced significantly with automation and improved detection methods. Innovations like thermocyclers, dUTP incorporation, and combined RT-PCR assays enhance efficiency and reduce errors in molecular diagnostics.

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Area of Science:

  • Molecular Biology
  • Biotechnology

Context:

  • Polymerase Chain Reaction (PCR) is a fundamental molecular biology technique.
  • Continuous advancements have been made since its inception to improve its efficiency, accuracy, and automation.

Purpose:

  • To review the key developmental highlights in PCR technology.
  • To discuss the progression towards automated PCR testing and its impact on molecular diagnostics.

Summary:

  • Early developments include the automation of amplification using thermocyclers.
  • Improvements such as dUTP incorporation with UNG (AmpErase) minimize false positives.
  • Combined reverse transcription PCR (RT-PCR) assays with single thermoactive enzymes (Tth DNA polymerase) simplify RNA amplification.
  • Modern methods for amplicon detection have eliminated radioactivity and electrophoresis, utilizing microwell plate assays for faster and simpler results.

Related Experiment Videos

  • The COBAS AMPLICOR system represents a major advancement, automating both amplification and detection, thereby reducing hands-on time and improving reproducibility.

    • Impact:

    • Automation in PCR, exemplified by systems like COBAS AMPLICOR, significantly reduces hands-on time and enhances assay reproducibility.
    • Ongoing developments, including kinetic PCR and 5' exonuclease assays, aim for further automation and efficiency in molecular testing.
    • These advancements are crucial for meeting the evolving demands of clinical requirements and molecular diagnostics.