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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

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Low-cost, real-time, continuous flow PCR system for pathogen detection.

B Leticia Fernández-Carballo^1,2, Ian McGuiness¹, Christine McBeth¹

¹Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.

Biomedical Microdevices

|March 21, 2016

Summary

This summary is machine-generated.

This study introduces a portable, low-cost Polymerase Chain Reaction (PCR) system for rapid pathogen detection. The continuous flow system enables faster results, making it ideal for point-of-care diagnostics.

Keywords:

Lab-on-a-chip Nucleic acid test On chip polymerase chain reaction (PCR)Pathogen diagnostic Point-of-care (POC)Real-time polymerase chain reaction (qPCR)

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High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

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Published on: November 28, 2016

Area of Science:

Biotechnology
Medical Diagnostics
Microbiology

Background:

Point-of-care (POC) diagnostics require rapid, accurate, and accessible pathogen detection methods.
Existing Polymerase Chain Reaction (PCR) technologies can be slow, expensive, and not easily deployable in resource-limited settings.

Purpose of the Study:

To develop and validate a portable, low-cost continuous flow PCR system for quantitative pathogen detection at the point-of-care.
To demonstrate the system's capability in detecting specific infectious bacteria, Chlamydia trachomatis and Escherichia coli O157:H7.

Main Methods:

A continuous flow PCR system was designed using a disposable thermoplastic chip manufactured via hot embossing for scalability.
The system incorporates two heaters, thermal sensors, and a real-time fluorescence optical detection system using FAM probes.
The system's performance was evaluated by determining the limit of detection, calculating PCR efficiencies, and testing various flow velocities for Chlamydia trachomatis and Escherichia coli O157:H7 detection.

Main Results:

The developed continuous flow PCR system demonstrated rapid amplification and detection of target pathogens.
Quantitative detection was achieved with a low limit of detection for both Chlamydia trachomatis and Escherichia coli O157:H7.
The system showed promising PCR efficiencies and versatility across different flow velocities.

Conclusions:

The portable, low-cost continuous flow PCR system offers a rapid and effective solution for point-of-care pathogen diagnostics.
The system's design, utilizing hot embossing and roll-to-roll compatibility, supports large-scale production and economic viability.
This technology holds significant potential for widespread application in infectious disease screening and monitoring.