Real-time Fluorescent Quantitative PCR

As an important tool in modern molecular biology, real-time fluorescent quantitative PCR technology has achieved remarkable achievements. However, it still faces some challenges in practical applications and also has broad development prospects.

The quality and processing of samples are the primary challenges faced by qPCR technology. The collection, preservation, and transportation of samples must be strictly standardized. Otherwise, it is very likely to lead to nucleic acid degradation or contamination, affecting the accuracy of the detection results. For example, if a blood sample is not separated into serum in a timely manner, the nucleases in it may degrade the nucleic acid; if an environmental sample is contaminated during the collection and transportation process, it will interfere with the detection results. Therefore, optimizing the sample processing procedure and developing more stable sample preservation methods are urgent problems to be solved.

The relatively high cost of instruments and reagents is also one of the factors limiting the widespread application of qPCR technology. qPCR instruments are expensive, and there are a wide variety of supporting reagents. Different detection items require different reagents, which makes the detection cost remain high. For some regions with limited resources or small laboratories, the high cost becomes an obstacle to its promotion and application. Reducing the cost of instruments and reagents and developing universal reagents are the keys to expanding the application scope of qPCR technology.

Technical errors are also issues that cannot be ignored. Errors in sample addition during the experimental operation process, slight differences in the reaction system, and errors in the detection of fluorescent signals, etc., may all lead to the repeatability and accuracy of the detection results being affected. In addition, the design of primers and probes also needs to be continuously optimized to meet different detection requirements and reduce problems such as non-specific amplification.

Despite the challenges, the future development prospects of qPCR technology are still very broad. With the continuous progress of technology, the instruments will become more miniaturized and portable, and the operation will also be more convenient. For example, the emergence of portable qPCR instruments makes it possible to apply them in on-site detection, primary medical care, and other scenarios, providing convenience for the prevention and control of infectious diseases and disease screening.

In terms of detection technology, qPCR technology will develop in combination with other technologies. Combining with microfluidic technology can achieve chip-based high-throughput and high-sensitivity detection; combining with next-generation sequencing technology can further improve the accuracy and comprehensiveness of detection and deeply explore gene information.

In the application field, qPCR technology will continue to expand. In addition to the existing fields of medical diagnosis and scientific research, it will also play an important role in more fields such as food safety, environmental monitoring, and agricultural breeding, making greater contributions to protecting human health and promoting social development.

Real-time fluorescent quantitative PCR technology faces challenges but is also full of development opportunities. By continuously solving existing problems and strengthening technological innovation, it will continue to shine in future technological development, providing more powerful technical support for humans to explore the mysteries of life and deal with various challenges.