The development of instruments that allow real-time monitoring of fluorescence within a PCR reaction vessel led to a significant advance in PCR technology and applications. Many different instruments and fluorescent probe systems have been developed and are currently available.

Real-time PCR assays can be completed rapidly since no manipulations are required post-amplification. Identification of the amplification products by probe detection in real-time is highly accurate compared with size analysis on gels. Analysis of the progress of the reaction allows accurate quantification of the target sequence over a very wide dynamic range, provided suitable standards are available. Further investigation of the real-time PCR products within the original reaction mixture using probes and melting analysis can detect sequence variants including single base mutations.

Real-time PCR has applications in many branches of biological science. Applications include gene expression analysis, the diagnosis of infectious disease and human genetic testing. Due to their fluorimetry capabilities, real-time machines are also compatible with alternative amplification methods such as NASBA, provided a fluorescence end-point is available.

Bibliography:

1. Real-Time PCR: Current Technology and Applications
2. Real-Time PCR in Microbiology: From Diagnosis to Characterization
3. PCR Books

from N.A. Saunders in Real-Time PCR: Current Technology and Applications

Labels: applications, assays, NASBA, probes, real-time pcr

It is clearly desirable to develop rapid and precise diagnostic PCR procedures for use in clinical laboratories. The authors of a recent review on the use of PCR for the diagnosis of malaria assert that PCR "should be considered as the gold standard for the diagnosis of imported malaria". Since the first description of the diagnosis of Plasmodium infection by PCR, the role of PCR in the laboratory diagnosis of imported malaria is still a topical question. However, the authors claim that PCR-based assays are more sensitive and more specific than all conventional methods. The highest contribution of the molecular diagnosis is that a negative PCR result would indicate the lack of any malaria infection, thus quickly orienting the investigations toward other aetiology.
from Berry et al. in Parasite 2008 15(3): 484-488

Bibliography:

1. Real-Time PCR in Microbiology: From Diagnosis to Characterization
2. Real-Time PCR: Current Technology and Applications
3. PCR Books
4. Malaria Parasites: Genomes and Molecular Biology

Labels: applications, clinical, diagnosis, microbiology