Real-time PCR for Mold Identification

The US EPA developed a detection system utilizing the TaqMan® Real-Time PCR system for detection and quantification of genetic material.

Essentially, this technology combines a fluorogenic element with that of traditional PCR.  In Q-PCR, we add a probe (a small oligonucleotide segment with bound fluorescent reporter and quencher), which is designed to hybridize to a DNA sequence on the target DNA between flanking primers.

During the annealing stage of amplification, the probe hybridizes to its sequence-specific complementary segment of target DNA. As the extension stage proceeds, the Taq polymerase functions to catalyze the addition of complementary nucleotides to the DNA backbone and, upon meeting with the probe cleaves, it releases the fluorophore (fluorescent reporter) from its quencher, allowing its excitation. This excitation is detected in real-time by the thermocycler.

Direct comparison of fluorescence detected in your sample to that of a standard curve of DNA amplification and fluorescence of a known quantity of spores allows Microbac to report sample concentration in number of spores per sampled material (i.e. ml, L, g).