Genetic PCR Solutions (GPS): The international transfer of PCR technology.
Detection of bacteria, virus, fungi & parasites through real time PCR analysis is a very fast, precise and sensitive technique that GPS now put within the reach of any laboratory. GPS selects specific equipment and high quality real time PCR testing kits available, suitable for any pathogen that you need to investigate. Our solutions can be applied to the analysis of food, water, air, pharmaceutical, cosmetics, veterinary and clinical research. GPS can design the PCR detection kit specific for the pathogen you propose and the new produced kit will be available for you in a few weeks. We provide a range of instruments selected for you specific needs: air sampler, tissue homogenizers, cell disruptors, DNA/RNA automated extractors, end-point and real time PCR thermo cyclers, etc. GPS ensuring that all of the products that our customers receive are meticulously quality controlled. Protocols using represented products have been validated by associated laboratories and international research institutions.
Real time PCR
Developed in the mid 1990 for the analysis and quantification of nucleic acids, real-time PCR has become the method of choice for molecular biological testing, gaining rapidly in popularity. It is based on the technique of the polymerase chain reaction (PCR) that was first envisioned by Kary Mullis almost 30 years ago, during a moonlit drive through the redwood hills of California. The technology of PCR has become one of the most influential discoveries of the molecular biology revolution and one for which Mullis received the Nobel Prize in 1993. Because of the impact of PCR and the thermostable Taq DNA polymerase (the enzyme responsible for the PCR revolution), the pair was named as the first “Molecule of the Year” by Science in 1989. In many ways, the recent development of real-time PCR seems set to change the general use of PCR. The advancement provided by the real-time version of PCR is due to its unique ability to monitor the complete DNA amplification process. During conventional PCR, the two strands of a DNA molecule are subjected to a series of heating and cooling cycles that result in DNA strand separation, oligonucleotide primer annealing, and thermos table Taq DNA polymerase– directed primer extension, ultimately generating two identical daughter strands. Iterative cycling of the process exponentially amplifies the number of original DNA molecules, hence the term PCR (Polymerase Chain Reaction). After completion of the PCR reaction, amplification products are analyzed by size fractionation of the amplified sample with the use of gel electrophoresis. In the mid 1990, researchers showed that the 5 nuclease activity of the Taq DNA polymerase could be exploited as a method to indirectly assess the level of DNA amplification with the use of specific fluorescent probes, eliminating the need for electrophoresis. Around the same time, researchers showed that real-time monitoring of the DNA amplification within the PCR reaction tube during the PCR could be achieved by using fluorescent DNA binding dyes, which is known as kinetic PCR (6). The coupling of these two processes led to today’s technology of fluorescence detection real-time PCR.
Microbial detection by real time PCR
Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has catalyzed wider acceptance of PCR because it is more rapid, sensitive and reproducible, while the risk of carryover contamination is minimized. Real-time PCR assays have been extremely useful for studying microbial agents of infectious disease, where they have helped to clarify many disease processes. Most of the assays presented in the literature have increased the frequency of microbial detection compared to non-PCR techniques, making the implementation of real-time PCR attractive to many. Of course, real-time PCR has also proven valuable for basic microbiological research, where its ability to amplify template from a wide array of sample types has made it an ideal system for application across the various microbiological disciplines.