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Updated: Feb 13, 2026

Serum and Plasma Copy Number Detection Using Real-time PCR

Serum and Plasma Copy Number Detection Using Real-time PCR

Published on: December 15, 2017

Real-time PCR detection chemistry.

E Navarro¹, G Serrano-Heras¹, M J Castaño¹

¹Research Unit, General University Hospital, Laurel s/n, 02006 Albacete, Spain.

Clinica Chimica Acta; International Journal of Clinical Chemistry

|December 3, 2014

Summary

This summary is machine-generated.

Real-time PCR offers sensitive and specific nucleic acid detection, making it ideal for diagnostics and food safety. This review categorizes fluorescent chemistries, including DNA intercalators and labeled probes, for advanced molecular analysis.

Keywords:

DNA binding dye DNA detection chemistries Fluorescent primer-probe Fluorescent probe Nucleic acid analogues Real-time PCR

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Area of Science:

Molecular Biology
Biochemistry
Analytical Chemistry

Background:

Real-time PCR is a preferred method for diagnostics and food applications due to its sensitivity, specificity, and reduced hands-on time.
It integrates polymerase chain reaction (PCR) chemistry with fluorescent reporters to monitor amplification in real-time, eliminating the need for post-PCR analysis.

Purpose of the Study:

To provide a comprehensive review of fluorescent-based nucleic acid analysis methods used in real-time PCR.
To classify and detail various real-time PCR chemistries and their applications.

Main Methods:

Classification of real-time PCR chemistries into two main groups: DNA intercalating agents and fluorophore-labeled oligonucleotides.
Sub-classification of fluorophore-labeled oligonucleotides into primer-probes, hydrolysis/hybridization probes, and nucleic acid analogues.
Detailed examination of structures, mechanisms, advantages, and applications of various fluorescent probes and analogues.

Main Results:

Real-time PCR chemistries are broadly divided into intercalating dyes (e.g., SYBR Green I, EvaGreen) and probe-based methods.
Probe-based methods are further categorized into primer-probes, hydrolysis probes (e.g., TaqMan), and hybridization probes (e.g., Molecular Beacons).
Nucleic acid analogues like PNA, LNA, and ZNA represent another significant category of real-time PCR detection molecules.

Conclusions:

Real-time PCR offers a powerful platform for sensitive and specific nucleic acid quantification.
The diversity of fluorescent chemistries, including intercalators, probes, and analogues, provides flexibility for various diagnostic and research applications.
Understanding these different fluorescent-based methods is crucial for optimizing real-time PCR assays.