FlashPCR: Revolutionising QPCR by Accelerating Amplification Through Low ∆T Protocols

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 Mar 13

PMID 38474020

Authors

Stephen A Bustin

Sara Kirvell

Tania Nolan

Gregory L Shipley

Affiliations

Soon will be listed here.

Abstract

Versatility, sensitivity, and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research, as well as diagnostic applications. However, for point-of-care (PoC) use, traditional qPCR faces two main challenges: long run times mean results are not available for half an hour or more, and the requisite high-temperature denaturation requires more robust and power-demanding instrumentation. This study addresses both issues and revises primer and probe designs, modified buffers, and low ∆T protocols which, together, speed up qPCR on conventional qPCR instruments and will allow for the development of robust, point-of-care devices. Our approach, called "FlashPCR", uses a protocol involving a 15-second denaturation at 79 °C, followed by repeated cycling for 1 s at 79 °C and 71 °C, together with high Tm primers and specific but simple buffers. It also allows for efficient reverse transcription as part of a one-step RT-qPCR protocol, making it universally applicable for both rapid research and diagnostic applications.

Citing Articles

Rapid Molecular Diagnostics of Pneumonia Caused by Gram-Negative Bacteria: A Clinician's Review.

Pintea-Simon I, Bancu L, Mare A, Ciurea C, Toma F, Man A Antibiotics (Basel). 2024; 13(9).

PMID: 39334980 PMC: 11429159. DOI: 10.3390/antibiotics13090805.

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