Real-Time PCR: History and Fluorogenic Chemistries
Ian M. Mackay, John F. Mackay, Michael D. Nissen, and Theo P. Sloots
from: Real-Time PCR in Microbiology: From Diagnosis to Characterization (Edited by: Ian M. Mackay). Caister Academic Press, U.K. (2007)
The use of real-time PCR in microbial diagnostics has increased to the point where it has evolved from a novelty into a mature and essential technology for the field. In doing so, real-time PCR has driven significant changes in the way we detect microbes. The predominantly phenotype-related methods of culture and antigen detection, while still of considerable value, are being supplanted by the detection, characterization and quantification of microbial nucleic acids. Real-time PCR has engendered wider acceptance of the PCR technique due to its improved rapidity, sensitivity, reproducibility and the considerably reduced risk of carry-over contamination. There are many fluorogenic chemistries that can detect PCR product as it accumulates but only a few are useful, popular or exciting enough to be the subject of publication in the field of microbiology. We review how real-time PCR has come to be; especially the essential role of fluorescence and we critically review the plethora of detection chemistries available to the end user read more ...