qfPCR (Quantitative Fluorescent PCR) is a technique that amplifies a specific DNA sequence and detects the amplified product with fluorescence. This technology is particularly used for rapid detection of copy number mutations of genes and chromosomal aberrations (e.g. trisomy). qfPCR amplifies DNA in the same way as regular PCR, but the amount of amplified DNA fragments can be quantitatively analyzed as a fluorescent signal.
Mechanism and Features of qfPCR
1. Use of fluorescent markers
- In qfPCR, primers labeled with fluorescent dyes are used to amplify the target DNA sequence. A fluorescent signal is generated according to the length and amount of the amplified product, and the amount of DNA is quantified based on the intensity of the fluorescence.
2. Quantitative Analysis
- The intensity of the fluorescent signal allows quantitative evaluation of DNA copy number and gene quantity. This allows for the detection of genetic or chromosomal abnormalities, if present, as a higher or lower copy number than normal.
3. Rapid Results
- Because qfPCR amplifies and detects only specific target regions in a short period of time, it provides results more rapidly than other genetic analysis methods. Therefore, it is important in prenatal diagnosis and detection of specific chromosomal abnormalities to be able to provide results quickly.
Main applications of qfPCR
- Detection of Chromosomal Aberrations
- qfPCR is often used to diagnose certain chromosomal abnormalities (e.g., trisomy). For example, it is used to detect trisomy of chromosomes 21 (Down syndrome), 18 (Edwards syndrome), and 13 (Patou syndrome).
- Prenatal Diagnosis
- Analysis of fetal DNA can be used to determine the presence or absence of chromosomal abnormalities. For example, qfPCR can be used to detect specific chromosome number abnormalities in samples collected during amniotic fluid and chorionic villus examinations to expedite prenatal diagnosis.
- Genetic copy number variation (CNV) analysis
- It is also used to evaluate whether a particular gene is over-copied or deleted. This allows detection of diseases and disorders associated with gene deletions and amplifications.
Conclusion
Quantitative Fluorescent PCR (qfPCR) is a technique that amplifies specific DNA sequences and quantitatively analyzes them by means of fluorescent signals. It is used in the detection of chromosomal abnormalities and prenatal diagnosis, and is characterized by its rapid results. For this reason, it is considered an important technology in the medical field to help diagnose genetic abnormalities and assess disease risk.
