AffiPCR® Aerobic complex Real Time PCR

Target Disease Type: Intestinal Infections

Specific Application:

Storage and Shipping : 4 weeks

Aerobic complex RT-PCR (CE) B88-100FRT DataSheet

NTENDED USE

Aerobic complex Real-TM Quant kit is a Real-Time test for the qualitative and quantitative detection of Enterobacteriaceae (E.coli, Klebsiella spp., Proteus spp. etc.), Staphylococcus spp. and Streptococcus spp. in biological materials by using real-time hybridization-fluorescence detection.

PRINCIPLE OF ASSAY
Detection and quantitation of Enterobacteriaceae, Staphylococcus spp. and Streptococcus spp. DNA by polymerase chain reaction (PCR) with real-time hybridization-fluorescence detection contains two steps: DNA extraction from the biological material and amplification of DNA fragment of microorganism with real-time hybridization-fluorescence detection. The DNA extraction from the clinical material is carried out with the presence of the Internal Control (Internal Control-FL), which allows controlling the procedure of examination of each sample. In the real-time PCR, the amplified product is detected with the use of fluorescent dyes. These dyes are linked to oligonucleotide probes, which bind specifically to the amplified product during thermocycling. The real-time monitoring of fluorescence intensities during the real-time PCR allows the detection of accumulating product without re-opening the reaction tubes after the PCR run. The results of amplification of Enterobacteriaceae, Staphylococcus spp. and Streptococcus spp. DNA are detected separately for each type by three different channels.

The DNA quantitation by real-time PCR is based on the existence of linear dependence between the logarithm of initial DNA-target concentration in the sample and start of theexponential growth of the fluorescent signal (threshold cycle, Ct) . For the quantitative analysis simultaneous carried out real-time amplification with real-time detection for the DNA samples obtained from the test samples and DNA-standards (the samples with the certain concentration of DNA-target) . According to the results of amplification DNA- standards built a calibration line on which the determination of the concentration of DNA-target in the test samples.